Therapeutic agents

ABSTRACT

A compound of formula (I) or a compound of formula (II) or pharmaceutically acceptable salts thereof, wherein R1-R7 and X are as defined in the description, and the use of these compounds in therapy, in particular in treating cancer or as an inhibitor of the interaction of the MDM2 protein with p53.

REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.13/001,372, filed 23 Dec. 2010, which is a U.S. National Stage under 35USC §371 of PCT/GB2009/001599, filed 25 Jun. 2009, which claims priorityto GB Application No. 0811643.6, filed 25 Jun. 2008. The entire contentsof each recited priority document is hereby specifically incorporated,in each of their respective entireties, for all purposes.

The invention relates to a series of isoindolin-1-one derivatives whichfind particular utility as pharmaceuticals, in particular in thetreatment of cancer.

Under conditions of stress such as hypoxia and DNA damage it is knownthat the cellular level of the protein p53 increases. P53 is known toinitiate transcription of a number of genes which govern progressionthrough the cell cycle, the initiation of DNA repair and programmed celldeath^(1,2). Thus, p53 is a tumour suppressor.

The activity of p53 is tightly regulated by the MDM2 protein, thetranscription of which is itself regulated by p53. p53 is inactivatedwhen it becomes bound to the p53 transactivation domain of the MDM2protein. Once inactivated the functions of p53 are repressed and thep53-MDM2 complex becomes a target for ubiquitinylation.

In normal cells the balance between active p53 and inactive MDM2-boundp53 is maintained in an autoregulatory negative feed back loop^(3,4).That is to say that p53 can activate MDM2 expression, which in turnleads to the repression of p53.

It has been found that inactivation of p53 by mutation is common inaround half of all tumours. Furthermore, in around 7% of tumours, overexpression of MDM2 results in the loss of functional p53, therebyallowing malignant transformation and uncontrolled tumour growths.

X-ray crystal studies of the MDM2-p53 complex have been conducted andhave revealed a hydrophobic pocket on the surface of MDM2 into which theside chains of Phe 19, Trp 23 and Leu 26 on p53 bind⁶. Therefore,inhibition of the MDM2-p53 binding interaction is an attractive targetfor researchers developing treatments for cancer as a means of restoringnormal p53 activity in cells overexpressing MDM2 and thereby exerting ananti-tumour effect.

A number of inhibitors of the MDM2-p53 interaction have been discoveredincluding peptide inhibitors, the natural product chlorofusion, andsmall molecules such as the imidazolines described in WO 03/051359⁸⁻¹¹.

The invention describes a novel series of compounds which inhibit theMDM2-p53 interaction and which have exciting in vitro activity.

According to a first aspect of the invention there is provided acompound of formula I:

or a compound of formula II:

or a pharmaceutically acceptable salt thereof, wherein in both formulaeI and II:

X is selected from O, N or S;

R¹ is selected from hydrogen, substituted or unsubstituted alkyl,substituted or unsubstituted hydroxyalkyl, substituted or unsubstitutedalkylamine, substituted or unsubstituted aryl, substituted orunsubstituted heteroaryl, substituted or unsubstituted aralkyl, andsubstituted or unsubstituted heteroaralkyl;

R² is selected from hydrogen, substituted or unsubstituted alkenyl oralkynyl, substituted or unsubstituted branched hydroxyalkyl, substitutedor unsubstituted cycloalkyl having 6 ring carbon atoms or greater,substituted or unsubstituted cycloalkenyl, hydroxyalkylaralkyl,hydroxyalkylheteroaralkyl, and a carboxylic acid-containing group;

R³ is selected from hydrogen, substituted or unsubstituted alkyl,substituted or unsubstituted hydroxyalkyl, substituted or unsubstitutedalkylamine, substituted or unsubstituted alkoxy, substituted orunsubstituted aryl, substituted or unsubstituted heteroaryl, substitutedor unsubstituted aralkyl, and substituted or unsubstitutedheteroaralkyl; and

R⁴-R⁷ represents groups R⁴, R⁵, R⁶ and R⁷ which are independentlyselected from hydrogen, halo, hydroxy, substituted or unsubstitutedalkyl, substituted or unsubstituted hydroxyalkyl, substituted orunsubstituted alkenyl, substituted or unsubstituted alkynyl, substitutedor unsubstituted heteroaryl, substituted or unsubstituted heteroaralkyl,substituted or unsubstituted alkylamine, substituted or unsubstitutedalkoxy, trifluoromethyl, amino, nitro, carboxyl, carbonyl,methylsulfone, trifluoromethylsulfone, cyano and substituted orunsubstituted sulfonamide;

wherein, where R² is substituted or unsubstituted branched hydroxyalkyl,X is O or S;

and wherein, where R² is hydrogen, at least one of R⁴-R⁷ is not hydrogenand R³ is not a benzimidazole derivative;

and wherein, in the formula II, the 6-membered ring may have 0, 1, or 2C═C double bonds.

According to a second aspect of the invention there is provided acompound of formula I or formula II, or a pharmaceutically acceptablesalt thereof, wherein

X is selected from O, N or S;

R¹ is selected from substituted aryl, substituted heteroaryl,substituted aralkyl, and substituted heteroaralkyl;

R² is selected from halo, acetyl, substituted or unsubstituted acyclicalkyl, substituted or unsubstituted hydroxyalkyl, substituted orunsubstituted alkylamine, substituted or unsubstituted alkoxyalkyl,substituted or unsubstituted aryl, substituted or unsubstitutedheteroaryl, substituted or unsubstituted aralkyl, and substituted orunsubstituted heteroalkyl;

R³ is selected from hydrogen, hydroxy, substituted or unsubstitutedalkyl, substituted or unsubstituted hydroxyalkyl, substituted orunsubstituted alkylamine, substituted or unsubstituted alkoxy,substituted or unsubstituted aryl or heteroaryl, and substituted orunsubstituted aralkyl or heteroalkyl; and

R⁴-R⁷ represents groups R⁴, R⁵, R⁶ and R⁷ which are independentlyselected from hydrogen, halo, hydroxy, substituted or unsubstitutedalkyl, substituted or unsubstituted hydroxyalkyl, substituted orunsubstituted alkenyl, substituted or unsubstituted alkynyl, substitutedor unsubstituted heteroaryl, substituted or unsubstituted heteroaralkyl,substituted or unsubstituted alkylamine, substituted or unsubstitutedalkoxy, trifluoromethyl, amino, nitro, carboxyl, carbonyl,methylsulfone, trifluoromethylsulfone, cyano and substituted orunsubstituted sulfonamide;

wherein when R² is a straight chain hydroxyalkyl, R¹ is not selectedfrom 4-nitrobenzyl or 4-chlorobenzyl;

and wherein, where R² is hydrogen, at least one of R⁴-R⁷ is not hydrogenand R³ is not a benzimidazole derivative or a benzimidazolinederivative;

and wherein when R³ is a phenyl group, R¹ cannot be 4-methoxybenzyl or4-hydroxybenzyl group;

and wherein, in the formula II, the 6-membered ring may have 0, 1, or 2C═C double bonds.

In the compounds of formula II, the 6-membered aromatic ring may becompletely saturated, or it may have one carbon-carbon double bond, oralternatively it may have two carbon-carbon double bonds. All of thesevariations are individually envisaged within the scope of the invention.

According to a third aspect of the invention there is provided acompound as defined in the first or second aspects for use in therapy.Further, in a fourth aspect of the invention there is provided acompound as defined in the first or second aspects for use in treatingcancer.

In a fifth aspect of the invention there is provided a compound asdefined in the first or second aspects, wherein said compound inhibitsthe interaction of MDM2 protein with p53.

In an sixth aspect of the invention there is provided a compound asdefined in the first or second aspects, for use as an activepharmaceutical substance for the treatment of cancer.

In an seventh aspect of the invention a compound of the first or secondaspects may be used in the manufacture of a medicament; and in an eighthaspect a compound of the first or second aspects may be used in themanufacture of a medicament for the treatment of cancer.

Also disclosed as a ninth aspect of the invention is a pharmaceuticalcomposition comprising an effective amount of at least one compound asdefined in the first or second aspects of the invention and apharmaceutically acceptable carrier.

In a tenth aspect of the invention there is provided a method oftreating a mammal comprising the steps of administering a medicamentcomprising at least one compound as defined in the first or secondaspects of the invention.

In an eleventh aspect of the invention there is provided a kitcomprising at least one compound as defined in the first or secondaspects of the invention; and instructions for use. The kit mayadditionally comprise a second compound as defined in the first orsecond aspects of the invention or an alternative cancer treatmentcompound known in the art.

Advantageously, the compounds of the present invention have been shownto be good inhibitors of the formation of the MDM2-p53 complex.

The term “halo” is used herein to denote a halogen atom which istypically selected from fluorine, chlorine, bromine or iodine.

The term “alkyl” is used herein to denote, in particular, a lower alkylgroup, i.e. a cyclic, branched (including ring structures formed via thelinking of two branches at the same carbon atom) or straight chainhydrocarbon having one to eight carbon atoms, for example propyl. Cyclicalkyls, or cycloalkyls are defined herein as non-aromatic saturatedhydrocarbons having at least one carbon-atom ring (typically having from6 to 10 ring carbon atoms), for example cyclohexyl or cyclooctyl.

The term “alkenyl” is used herein to denote an alkyl group including oneor more carbon-carbon double bonds, for example butenyl orcyclopentenyl.

The term “alkynyl” is used herein to denote an alkyl group including oneor more carbon-carbon triple bonds, for example butynyl.

The term “aryl” is used herein to denote a carbocyclic group orstructure having at least one aromatic ring. The said ring may form partof a multiple condensed ring structure, for example phenyl, naphthaleneor anthracene.

The term “aralkyl” is used herein to denote an alkyl chain, ashereinbefore defined, in which there is an aryl group attached thereto,as hereinbefore defined, for example benzyl.

The term “heteroaryl” is used herein the denote an aryl group, ashereinbefore defined in which said group comprises at least oneheteroatom, selected from, for example N, O or S, in said at least onearomatic ring. Examples of heteroaryl groups which may be used inaccordance with the invention include, but are not limited to, pyridine,pyrrole, furan, thiophene and imidazole.

The term “heteroaralkyl” is used herein to denote an aralkylsubstituent, as hereinbefore defined, in which said at least onearomatic ring comprises at least one heteroatom selected from, forexample N, O or S. Examples of heteroaralkyl groups which may be used inaccordance with the invention include, but are not limited to,methylpyridine and methylfuran.

The term “substituted alkyl” is used herein to denote an alkylsubstituent, as hereinbefore defined, which is substituted with one ormore functional groups.

The term “substituted alkenyl” is used herein to denote an alkenylsubstituent, as herein before defined, which is substituted with one ormore functional groups.

The term “substituted alkynyl” is used herein to denote an alkynylsubstituent, as hereinbefore defined, which is substituted with one ormore functional groups.

The term “substituted aryl” is used herein to denote an arylsubstituent, as hereinbefore defined, which is substituted with one ormore functional groups. Examples of substituted aryl groups which may beused in accordance with the invention include, but are not limited to,halophenyl, methylphenyl, nitrophenyl or cyanophenyl.

The term “substituted heteroaryl” is used herein to denote a heteroarylsubstituent, as hereinbefore defined, which is substituted with one ormore functional groups.

The term “substituted aralkyl” is used herein to denote an aralkylsubstituent, as hereinbefore defined, which is substituted with one ormore functional groups. Examples of substituted aralkyl groups which maybe used in accordance with the invention include, but are not limitedto, halobenzyl, benzonitrile, acetylbenzyl, benzoylbenzyl, nitrobenzyl,cyanobenzyl, methoxybenzyl, carboxamidobenzyl, or methylbenzyl.

The term “substituted heteroaralkyl” is used herein to denote aheteroaralkyl substituent, as hereinbefore defined, which is substitutedwith one or more functional groups.

The term “alkoxy” is used herein to denote an alkyl group, ashereinbefore defined, which is linked to a second chemical structure,which may be any of the foregoing, by way of an oxygen atom. The carbonchain of the alkyl group may be substituted with one or more functionalgroups to provide a “substituted alkoxy”. Examples of alkoxy groupswhich may be used in accordance with the invention include, but are notlimited to, ethoxy, methoxy and propoxy.

The term “alkylamine” is used herein to denote an alkyl group, ashereinbefore defined, comprising at least one amine function. The carbonchain of the alkyl group may be substituted with one or more functionalgroups. The amine function may be primary, secondary or tertiary.Examples of alkylamine groups which may be used in accordance with theinvention include, but are not limited to, ethylamine and diethylamine.The amine function may form part of a cyclic or heteroaromatic structureor another functionality, for example amide.

As referred to herein suitable functional groups for substitution asdescribed above include, but are not limited to, any of the followingwhich may be used alone or in combination: halo, hydroxyl, hydroxyalkyl,acyl, acetamide, carboxyl, cyano, carboxamide (carbamoyl), sulfonamide,sulfone, sulfoxide, amino, alkoxy or silico ligand.

Compounds of interest include those of formula I or formula II asdefined in the first aspect of the invention wherein R¹ is selected fromsubstituted or unsubstituted aryl, and substituted or unsubstitutedaralkyl; R² is selected from hydroxyalkenyl, hydroxyalkynyl, branched5-carbon hydroxyalkyl, hydroxycycloalkyl, hydroxycycloalkenyl,hydroxymethylcycloalkyl, hydroxymethylcycloalkylmethylene, andhydroxylalkylbenzyl; and R³ is selected from substituted orunsubstituted aryl, and substituted or unsubstituted aralkyl.

Where R¹ is selected from substituted or unsubstituted aryl, andsubstituted or unsubstituted aralkyl it is typically a substitutedaralkyl, particularly a substituted benzyl. The substituted benzyl maybe, for instance, benzonitrile, chlorobenzyl, bromobenzyl, iodobenzyl,methylbenzyl, acetylbenzyl, benzoylbenzyl, cyanobenzyl, methoxybenzyl,carboxamidobenzyl, or nitrobenzyl. Typically, the substituted benzyl isnitrobenzyl, however isosteres of this functional group may also beadvantageously used such as benzoyl, benzylcarboxylate,benzylcarboxylate alkyl ester, benzylsulfoxide or benzylsulfone. Othersubstituted benzyl groups may also be used, such as bromobenzyl,iodobenzyl, azobenzyl, aminobenzyl, or thioetherbenzyls.

The substituted benzyl may be substituted with one or more functionalgroups at any of positions 2- to 6-, however it is typical that a singlesubstituent is present at the 3-, 4- or 5-position, typically the4-position. Alternatively, R¹ may be a propyl group or unsubstitutedbenzyl.

Where R¹ is selected from substituted or unsubstituted heteroaryl,nitrogen is typically the heteroatom in the ring, and the heteroaryl istypically an unsubstituted pyridyl. Alternatively, the heteroaryl couldbe an N-pyridine oxide. In both embodiments, the nitrogen atom may be inthe 2-, 3-, or 4-position.

In a further embodiment of R¹, there may be a further alkylene groupbetween the benzyl moiety and the nitrogen atom of the isoindolinone.Alternatively, the alkyl carbon atom of the benzyl may be furthersubstituted with an alkyl group, such as a methyl group.

Typically, when R² is an alkenyl substituent, it will be selected fromhydroxybutenyl, hydroxycyclohexenyl or hydroxycyclopentenyl, most often4-hydroxybut-2-enyl, 4-hydroxycyclohex-2-enyl or4-hydroxycyclopent-2-enyl. However, when R² is a hydroxyalkynyl, it istypically a hydroxybutynyl, typically 4-hydroxybut-2-ynyl.

When R² is a branched 5-carbon hydroxyalkyl, it is typically a branchedpropyl chain such as hydroxy-2,2-dimethylpropyl orhydroxy-2,2-cyclopropylpropyl. The hydroxyl group will typically be aterminal hydroxy as would be found in 3-hydroxy-2,2-dimethylpropyl or1-hydroxy-2,2-cyclopropylpropyl.

In embodiments where R² is a substituted or unsubstituted cycloalkylhaving 6 ring carbon atoms or greater it will typically be selected fromhydroxycyclooctyl, hydroxymethylcyclohexylmethylene, andhydroxycyclohexyl. Typically the selection will be from5-hydroxycyclooctyl, 2-hydroxymethylcyclohexylmethylene,4-hydroxycyclohexyl, and 4-hydroxymethylcyclohexylmethylene.

In alternative embodiments R² may be a hydroxyalkylbenzyl, it is typicalin these embodiments that R² is a hydroxymethylbenzyl such as4-hydroxymethylbenzyl or 3-hydroxymethylbenzyl. Alternatively, R² maycomprise a linear alkoxy or amino group, such as an alkylamine wherealkyl has the same meaning as given hereinabove. Typical amines includen-propylamine.

According to a further embodiment of the invention, when R² is acarboxylic acid-containing group, it may be a succinic acid-containinggroup, such as a succinic acid methyl cyclopropylmethyl group.

According to a further embodiment of the invention, R² may be a2-hydroxymethyl allyl group.

According to a further embodiment of the invention, R² may be hydrogenwhere at least one of R⁴-R⁷ is not hydrogen and R³ is not abenzimidazole derivative.

Typically, R³ be selected from a singly substituted phenyl, typically ahalophenyl, often chlorophenyl or bromophenyl, most often4-chlorophenyl, although the substituent (regardless of identity) may beat any of the 2- to 6-positions.

Additional compounds of interest include those of formula I or formulaII as defined in the second aspect of the invention wherein R¹ issubstituted aralkyl, R² is acyclic hydroxyalkyl, and R³ is substitutedaryl. In these embodiments R¹, may be any of the substituents describedabove with regard to the first aspect of the invention, however it istypical that R¹ is a substituted 1-ethylphenyl or a substituted benzyl.

Although there may be more than one substituent which may be substitutedat any of the 2- to 6-positions on the encompassed phenyl group, it istypical that only a single substituent be present, often a halo group,typically present at the 4-position. Often the halophenyl will be achlorophenyl and it is currently typical that when R¹ is a substituted1-ethylphenyl, R¹ is 1-(4-chlorophenyl)-ethyl. Either the R— or theS-enantiomer of this substituted 1-ethylphenyl substituent may be used,however the S-enantiomer is typical.

When R¹ is a substituted benzyl, it is typically nitrobenzyl, moretypically 4-nitrobenzyl, 4-cyanobenzyl, 4-chlorobenzyl, 4-bromobenzyl,or 4-iodobenzyl. Most typically, R¹ is 4-nitrobenzyl.

Where R² is acyclic hydroxyalkyl it is typical that the alkyl is ann-alkyl chain. Typically, the chain length is 2 to 6 carbons, typically4 carbons. The most typical acyclic hydroxyalkyl is 4-hydroxy-n-butyl orhydroxypropyl.

As with the first aspect of the invention it is typical that R³ isselected from a singly substituted phenyl, typically a halophenyl, oftenchlorophenyl or bromophenyl, most often 4-chlorophenyl, although thesubstituent (regardless of identity) may be at any of the 2- to6-positions.

In typical embodiments of the compounds of the first and second aspectsof the invention, R⁴, R⁵, R⁶ and R⁷ are each hydrogen atoms and X isindependently 0. However, in some compounds, one or more of R⁴ to R⁷ isH with two of the remaining R groups linked so as to form a 5- to7-membered ring structure. The ring structure is typically saturated andmay comprise at least one heteroatom selected from N, O or S.Alternatively, R⁴ to R⁷ may each be independently selected from methyl,t-butyl, chlorine, bromine and fluorine. In some aspects of theinvention, one (or more) of R⁴-R⁷ may independently be chlorine, or oneof R⁴-R⁷ may be chlorine and another may be fluorine.

It will be understood that where reference is made in this specificationto compounds of formulae I or II such reference should be construed asextending also to their pharmaceutically acceptable salts and to otherpharmaceutically acceptable bio precursors (for instance, prodrug,chemically protected or solvated forms) where relevant.

Salts

The term “salt” is used in the specification to denote the combinationof a charged form of a compound with an oppositely charged ion toproduce a neutral product. Examples of pharmaceutically acceptable saltsare discussed in Berge et al., 1977, “Pharmaceutically AcceptableSalts.” J. Pharm. ScL. Vol. 66, pp. 1-19.

Unless otherwise specified, a reference to a particular compound alsoincludes salt forms thereof.

For example, if the compound is anionic, or has a functional group whichmay be anionic (such as, —COOH may be —COO⁻), then a salt may be formedwith a suitable cation. Examples of suitable inorganic cations include,but are not limited to, alkali metal ions such as Na⁺ and K⁺, alkalineearth cations such as Ca²⁺ and Mg²⁺, and other cations such as Al³⁺.Examples of suitable organic cations include, but are not limited to,ammonium ion (i.e., NH₄ ⁺) and substituted ammonium ions (for example,NH₃R⁺, NH₂R₂ ⁺, NHR₃ ⁺, NR₄ ⁺). Examples of some suitable substitutedammonium ions are those derived from: ethylamine, diethylamine,dicyclohexylamine, triethylamine, butylamine, ethylenediamine,ethanolamine, diethanolamine, piperazine, benzylamine,phenylbenzylamine, choline, meglumine, and tromethamine, as well asamino acids, such as lysine and arginine. An example of a commonquaternary ammonium ion is N(CH₃)₄ ⁺.

If the compound is cationic, or has a functional group that may becationic (such as, —NH₂ may be —NH₃ ⁺), then a salt may be formed with asuitable anion. Examples of suitable inorganic anions include, but arenot limited to, those derived from the following inorganic acids:hydrochloric, hydrobromic, hydroiodic, sulfuric, sulfurous, nitric,nitrous, phosphoric, and phosphorous.

Examples of suitable organic anions include, but are not limited to,those derived from the following organic acids: 2-acetyoxybenzoic,acetic, ascorbic, aspartic, benzoic, camphorsulfonic, cinnamic, citric,edetic, ethanedisulfonic, ethanesulfonic, fumaric, glucheptonic,gluconic, glutamic, glycolic, hydroxymaleic, hydroxynaphthalenecarboxylic, isethionic, lactic, lactobionic, lauric, maleic, malic,methanesulfonic, mucic, oleic, oxalic, palmitic, pamoic, pantothenic,phenylacetic, phenylsulfonic, propionic, pyruvic, salicylic, stearic,succinic, sulfanilic, tartaric, toluenesulfonic, and valeric. Examplesof suitable polymeric organic anions include, but are not limited to,those derived from the following polymeric acids: tannic acid,carboxymethyl cellulose.

Prodrugs

The term “prodrug” is used in the specification to denote modified formsor derivatives of a pharmacologically active compound which biodegradeor are modified in vivo so as to become converted into said activecompound after administration, especially intravenous administration, inthe course of therapeutic treatment of a mammal. Typically, the prodrugis inactive, or less active than the active compound, but may provideadvantageous handling, administration, or metabolic properties. Suchprodrugs are commonly chosen because of an enhanced solubility ofaqueous media which helps to overcome formulation problems, and also insome cases to give a relatively slow or controlled release of the activeagent.

Unless otherwise specified, a reference to a particular compound alsoincludes prodrugs thereof.

For example, some prodrugs are esters of the active compound (forinstance, a physiologically acceptable metabolically labile ester).During metabolism, the ester group (—C(═O)OR) is cleaved to yield theactive drug. Such esters may be formed by esterification, for example,of any of the carboxylic acid groups (—C(═O)OH) in the parent compound,with, where appropriate, prior protection of any other reactive groupspresent in the parent compound, followed by deprotection if required.

Also, some prodrugs are activated enzymatically to yield the activecompound, or a compound which, upon further chemical reaction, yieldsthe active compound (for example, as in ADEPT, GDEPT or LIDEPT). Forexample, the prodrug may be a sugar derivative or other glycosideconjugate, or may be an amino acid ester derivative.

Solvates

The term “solvate” is used in the specification to denote a complex ofsolute (for instance the active compound or salt of active compound) andsolvent. If the solvent is water, the solvate may be convenientlyreferred to as a hydrate, for example, a mono-hydrate, a di-hydrate, atri-hydrate, etc.

Unless otherwise specified, a reference to a particular compound alsoincludes solvate forms thereof.

Chemically Protected Forms

The term “chemically protected form” is used in the specification todenote a compound in which one or more reactive functional groups areprotected from undesirable chemical reactions under specified conditions(for instance, pH, temperature, radiation, solvent, and the like). Inpractice, well known chemical methods are employed to reversibly renderunreactive a functional group, which otherwise would be reactive, underspecified conditions. In a chemically protected form, one or morereactive functional groups are in the form of a protected or protectinggroup (also known as a masked or masking group or a blocked or blockinggroup). By protecting a reactive functional group, reactions involvingother unprotected reactive functional groups can be performed, withoutaffecting the protected group; the protecting group may be removed,usually in a subsequent step, without substantially affecting theremainder of the molecule.

Unless otherwise specified, a reference to a particular compound alsoincludes chemically protected forms thereof.

A wide variety of such “protecting,” “blocking,” or “masking” methodsare used and well known in organic synthesis. For example, a compoundwhich has two non-equivalent reactive functional groups, both of whichwould be reactive under specified conditions, may be derivatized torender one of the functional groups “protected,” and thereforeunreactive, under the specified conditions; so protected, the compoundmay be used as a reactant which has effectively only one reactivefunctional group. After the desired reaction (involving the otherfunctional group) is complete, the protected group may be “deprotected”to return it to its original functionality.

For example, a hydroxy group may be protected as an ether (—OR) or anester (—OC(═O)R), for example, as: a t-butyl ether; a benzyl, benzhydryl(diphenylmethyl), or trityl (triphenylmethyl)ether; a trimethylsilyl ort-butyldimethylsilyl ether; or an acetyl ester (—OC(═O)CH₃, —OAc).

It should be understood that all plausible and compatible combinationsof the embodiments described above are explicitly disclosed herein. Eachof these combinations is disclosed herein to the same extent as if eachindividual combination was specifically and individually recited. Itshould also be understood that where any of the compounds referred tocan exist in one or more particular geometric, optical, enantiomeric,diasteriomeric, epimeric, atropic, stereoisomeric, tautomeric,conformational, or anomeric forms, including but not limited to, cis-and trans-forms; E- and Z-forms; c-, t-, and r-forms; endo- andexo-forms; R—, S—, and meso-forms; D- and L-forms; d- and l-forms; (+)and (−) forms; keto-, enol-, and enolate-forms; syn- and anti-forms;synclinal- and anticlinal-forms; α- and β-forms; axial and equatorialforms; boat-, chair-, twist-, envelope-, and half chair-forms; andcombinations thereof, hereinafter collectively referred to as “isomers”(or “isomeric forms”) all such forms, mixtures thereof, and theirpreparation and uses are within the scope of the invention.

It should be noted, however, that stereo chemical considerations arelikely to be important and there may be considerable selectivity suchthat different enantiomers or diastereoisomers have significantlydifferent inhibitory activity.

Examples of isoindolin-1-one compounds which are at present ofparticular interest or typically of use in carrying out the inventioncomprise the following:

TABLE 1 ELISA IC₅₀ Number Compound Name Structure (μM)* NU82923-(4-Chlorophenyl)-3-(4- hydroxycyclopent-2- enyloxy)-2-(4-(aminomethyl) benzonitrile)isoindolin-1- one

2.3 ± 0.3 NU8293 3-(4-Chlorophenyl)-3-(4- hydroxycyclopent-2-enyloxy)-2-(4- chlorophenyl)isoindolin- 1-one

1.5 ± 0.3 NU8294 3-(4-chlorophenyl)-3-(4- hydroxycyclopentenyloxy)-2-propylisoindolin-1- one 389.13 C₂₂H₂₂ClNO₃

15.1 ± 2.1 NU8295 3-(4-Chlorophenyl)-3-(4- hydroxycyclopent-2-enyloxy)-2-(4- methylbenzyl)isoindolin- 1-one

1.4 ± 0.4 NU8298 2-benzyl-3-(4- chlorophenyl)-3- ((1R,4S)-4-hydroxycyclopent-2- enyloxy)isoindolin-1-one

4.7 ± 1.1 NU8362 3-(4-Chlorophenyl)-3-(4- hydroxybutoxy)-2-[2-(4-nitrophenyl)-ethyl]-2,3- dihydroisoindol-1-one

3.2 ± 1.0 NU8368 3-(4-Chlorophenyl)-2-[1- (4-chlorophenyl)-ethyl]-3-(4-hydroxybutoxy)- 2,3-dihydro-isoindol-1- one

2.5 ± 0.8 NU8370 4-[1-(4-Chlorophenyl)-1- (4-hydroxybutoxy)-3-oxo-1,3-dihydro- isoindol-2-ylmethyl]- benzonitrile

3.5 ± 1.5 *Mean ± SE of n = 3 repeat experiments unless otherwiseindicated

Particularly useful examples of isoindolin-1-ones for use in carryingout the invention and which have been found to have particularly potentactivity comprise the following:

TABLE 2 ELISA IC₅₀ Number Compound Name Structure (nM)* NU82973-(4-Chlorophenyl)-3-(4- hydroxycyclopent-2- enyloxy)-2-(4-nitrobenzyl)isoindolin-1- one

1.4 ± 0.3 μM (n = 3) NU8350 3-(4-Chlorophenyl)-3-(4-hydroxybut-2-enyloxy)-2- (4-nitrobenzyl)-2,3- dihydroisoindol-1-one

402 ± 9  NU8351 3-(4-Chlorophenyl)-3-(4- hydroxybut-2-enyloxy)-2-(4-nitrobenzyl)-2,3- dihydroisoindol-1-one

405 ± 10  NU8352 3-(4-Chlorophenyl)-3-(5- hydroxycyclooctyloxy)-2-(4-nitrobenzyl)-2,3- dihydroisoindol-1-one

375 ± 36  (n = 5) NU8353 3-(4-Chlorophenyl)-3-(3- hydroxy-2,2-dimethylpropoxy)-2-(4- nitrobenzyl)-2,3-dihydro- isoindol-1-one

395 ± 75  NU8354 3-(4-Chlorophenyl)-3-(1- hydroxymethylcyclopropylmethoxy)-2-(4- nitrobenzyl)-2,3- dihydroisoindol-1-one

298 ± 22  (n = 8) NU8354A (+)-3-(4-Chlorophenyl)- 3-(1-hydroxymethylcyclopropyl methoxy)-2-(4-nitro- benzyl)-2,3-dihydroisoindol-1-one 164 ± 18  (n = 7) NU8354B (−)-3-(4-Chlorophenyl)-3-(1- hydroxymethylcyclopropyl methoxy)-2-(4- nitrobenzyl)-2,3-dihydroisoindol-1-one 1333 ± 120  (n = 7) NU83573-(4-Chlorophenyl)-3-(4- hydroxybut-2-ynyloxy)-2- (4-nitrobenzyl)-2,3-dihydroisoindol-1-one

656 ± 113 (n = 5) NU8358 3-(4-Chlorophenyl)-3-(4-hydroxymethylcyclohexyl methoxy)-2-(4- nitrobenzyl)-2,3-dihydroisoindol-1-one

582 ± 75  (n = 5) NU8359 3-(4-Chlorophenyl)-3-(2-hydroxymethylcyclohexyl methoxy)-2-(4- nitrobenzyl)-2,3-dihydroisoindol-1-one

569 ± 74  (n = 5) NU8360 3-(4-Chlorophenyl)-3-(4-hydroxycyclohexyloxy)-2- (4-nitrobenzyl)-2,3- dihydroisoindol-1-one

388 ± 107 (n = 5) NU8361 3-(4-Chlorophenyl)-3-(4- hydroxy-cyclohex-2-enyloxy)-2-(4-nitrobenzyl)- 2,3-dihydroisoindol-1-one

306 ± 92  (n = 5) NU8365 3-(4-Chlorophenyl)-2-[1-(4-chlorophenyl)-ethyl]-3- (4-hydroxybutoxy)-2,3- dihydroisoindol-1-one

869 ± 64  NU8366 3-(4-Chlorophenyl)-3-(4- hydroxymethylbenzyloxy)-2-(4-nitro-benzyl)-2,3- dihydroisoindol-1-one

983 ± 170 NU8367 3-(4-Chlorophenyl)-3-(3- hydroxymethylbenzyloxy)-2-(4-nitrobenzyl)-2,3- dihydroisoindol-1-one

732 ± 126 *Mean ± SE of n = 3 repeat experiments unless otherwiseindicated

Further typical examples of isoindolin-1-ones for use in carrying outthe invention and which have been found to have particularly potentactivity comprise the following:

TABLE 3 MDM2 IC₅₀ Number Compound Descriptions Structure (μm) NU83803-(3-aminopropoxy)-3- (4-chlorophenyl)-2-(4- nitrobenzyl)isoindolin-1-one

2.46 ± 0.39 μM (n = 5) NU8390 3-(4-bromophenyl)-3- (4-hydroxybutoxy)-2-(4-nitrobenzyl)isoindolin-1-one

570 ± 59  nM NU8391 3-(4-bromophenyl)-3-((1- (hydroxymethyl)cyclopropyl)methoxy)-2-(4-nitrobenzyl) isoindolin-1-one

368 ± 45  nM NU8392 3-(4-bromophenyl)-3-(3- hydroxypropoxy)-2-(4-nitrobenzyl)isoindolin-1-one

 1.4 ± 0.16 μM NU8393 3-(4-chlorophenyl)-3- hydroxy-4-methyl-2-(4-nitrobenzyl)isoindolin-1-one

 2.2 ± 0.34 μM NU8394 3-(4-chlorophenyl)-3- hydroxy-7-methyl-2-(4-nitrobenzyl)isoindolin-1-one

6.9 ± 2.7 μM NU8395 3-(4-chlorophenyl)-3- hydroxy-5-methyl-2-(4-nitrobenzyl)isoindolin-1-one

5.08 ± 0.65 μM NU8396 5-tert-butyl-3-(4-chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl) isoindolin-1-one

12.7 ± 1.4  μM NU8397 6-tert-butyl-3-(4-chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl) isoindolin-1-one

837 ± 49  nM NU8398 4-chloro-3-(4-chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl) isoindolin-1-one

510 ± 32  nM NU8399 6-tert-butyl-3-(4-chlorophenyl)-3-((1-(hydroxymethyl) cyclopropyl)methoxy)-2-(4-nitrobenzyl)isoindolin-1-one

152 ± 27  nM NCL- 000104 85 3-(4-chlorophenyl)-5-fluoro-3-hydroxy-2-(4-nitrobenzyl) isoindolin-1-one

3.76 ± 0.54 NCL- 000104 86 3-(4-chlorophenyl)-6-fluoro-3-hydroxy-2-(4-nitrobenzyl) isoindolin-1-one

5.19 ± 1.51 NCL- 000104 87 5,6-dichloro-3-(4- chlorophenyl)-3-((1-(hydroxymethyl)cyclopropyl) methoxy)-2-(4- nitrobenzyl)isoindolin-1-one

3.67 ± 1.15 NCL- 000104 88 4-((7-chloro-1-(4-chlorophenyl)-1-hydroxy-3-oxoisoindolin-2-yl) methyl)benzonitrile

1.62 ± 0.97 NCL- 000104 89 4-((4-chloro-1-(4-chlorophenyl)-1-hydroxy-3-oxoisoindolin-2-yl) methyl)benzonitrile

8.95 ± 2.16 NCL- 000104 90 2-(4-bromobenzyl)-4-chloro-3-(4-chlorophenyl)-3-hydroxyisoindolin- 1-one

0.847 ± 0.082 NCL- 000104 92 4-((7-chloro-1-(4-chlorophenyl)-1-((1-(hydroxymethyl)cyclopropyl) methoxy)-3-oxoisoindolin-2-yl)methyl)benzonitrile

0.185 ± 0.017 NCL- 000104 93 2-(4-bromobenzyl)-4-chloro-3-(4-chlorophenyl)-3-((1-(hydroxymethyl) cyclopropyl)methoxy)isoindolin-1-one

0.169 ± 0.003 NCL- 000104 94 3-(4-chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl)-2,3,4,5,6,7- hexahydro-1H-isoindol-1-one

2.81 ± 0.07 NCL- 000104 95 3-(4-chlorophenyl)-5-fluoro-3-((1-(hydroxymethyl)cyclopropyl) methoxy)-2-(4-nitrobenzyl) isoindolin-1-one

0.295 ± 0.065 NCL- 000104 96 3-(4-chlorophenyl)-6-fluoro-3-((1-(hydroxymethyl)cyclopropyl) methoxy)-2-(4-nitrobenzyl) isoindolin-1-one

0.852 ± 0.09 

TABLE 4 MDM2 Number Compound Description Structure IC₅₀ (μM) NU84005-tert-butyl-3-(4- chlorophenyl)-3- ((1-(hydroxymethyl)cyclopropyl)methoxy)- 2-(4-nitrobenzyl) isoindolin-1-one

733 ± 29  nM NU8401 3-(4-chlorophenyl)-3- ((1-(hydroxymethyl)cyclopropyl)methoxy)- 5-methyl-2-(4- nitrobenzyl) isoindolin-1-one

492 ± 35  nM NU8405 3-(4-chlorophenyl)-3- ((1-(hydroxymethyl)cyclopropyl)methoxy)- 4-methyl-2-(4- nitrobenzyl) isoindolin-1-one

274 ± 35  nM NU8406 4-chloro-3-(4- chlorophenyl)-3- ((1-(hydroxymethyl)cyclopropyl)methoxy)- 2-(4-nitrobenzyl) isoindolin-1- one

143 ± 26  nM NU8412 3-(4-chlorophenyl)-3- hydroxy-6-methyl-2-(4-nitrobenzyl) isoindolin-1-one

1.46 ± 0.36 NU8413 6-bromo-3-(4- chlorophenyl)-3- hydroxy-2-(4-nitrobenzyl) isoindolin-1-one

5.37 ± 0.51 NU8414 5-bromo-3-(4- chlorophenyl)-3- hydroxy-2-(4-nitrobenzyl) isoindolin-1-one

5.68 ± 0.16 NU8415 4-((1-(4-chlorophenyl)- 1-((1-(hydroxymethyl)cyclopropyl)methoxy)- 3-oxoisoindolin-2- yl)methyl)benzonitrile

1.79 ± 0.67 NU8416 2-(4-chlorobenzyl)-3-(4- chlorophenyl)-3-((1-(hydroxymethyl) cyclopropyl)methoxy) isoindolin-1-one

2.31 ± 0.56 NU8417 2-(4-bromobenzyl)- 3-(4-chlorophenyl)-3-((1-(hydroxymethyl) cyclopropyl)methoxy) isoindolin-1-one

1.20 ± 0.61 NU8418 3-(4-chlorophenyl)- 2-((R)-1-(4- chlorophenyl)ethyl)-3-((1- (hydroxymethyl) cyclopropyl)methoxy) isoindolin-1-one

8.8 ± 2.1 NU8419 3-(4-chlorophenyl)- 2-((S)-1-(4- chlorophenyl)ethyl)-3-((1- (hydroxymethyl) cyclopropyl)methoxy) isoindolin-1-one

8.9 ± 1.9 NU8424 5-bromo-3-(4- chlorophenyl)-3-((1- (hydroxymethyl)cyclopropyl)methoxy)- 2-(4-nitrobenzyl) isoindolin-1-one

902 ± 71  nM NU8425 6-bromo-3-(4- chlorophenyl)-3-((1- (hydroxymethyl)cyclopropyl)methoxy)- 2-(4-nitrobenzyl) isoindolin-1-one

1.03 ± 0.04 NU8429 3-(4-chlorophenyl)-3- ((1-(hydroxymethyl)cyclopropyl)methoxy)- 2-(pyridin-2-ylmethyl) isoindolin-1-one

10.6 ± 0.8 

TABLE 5 MDM2-p53 ELISA Name Structure IC₅₀ (uM) NU8398 4-chloro-3-(4-chlorophenyl)-3- hydroxy-2-(4- nitrobenzyl) isoindolin-1-one

0.51 ± 0.03 NCL-00013774 (?)-4-chloro-3-(4- chlorophenyl)-3-((1-(hydroxymethyl) cyclopropyl) methoxy)-2-(4- nitrobenzyl)isoindolin-1-one

 0.04 ± 0.004 NCL-00013775 (?)-4-chloro-3-(4- chlorophenyl)-3-((1-(hydroxymethyl) cyclopropyl) methoxy)-2-(4- nitrobenzyl)isoindolin-1-one

 1.26 ± 0.008 NCL-00016654 7-chloro-3-(4- chloro-phenyl)-3-hydroxy-2-(4- nitro-benzyl)-2,3- dihydro- isoindol-1-one

4.54  NCL-00016045 2-(4-acetylbenzyl)- 3-(4-chlorophenyl)-3-hydroxy-2,3- dihydroisoindol- 1-one

89   NCL-00014529 3-(4-chlorophenyl)- 3-(1-hydroxymethyl-cyclopropylmethoxy)- 2-(4-iodobenzyl)- 2,3-dihydroisoindol- 1-one

1.5  NCL-00014531 2-(4-acetylbenzyl)- 3-(4-chlorophenyl)- 3-(1-hydroxy-methylcyclopropyl- methoxy)-2,3- dihydroisoindol-1-one

17   NCL-00016046 3-(4-chlorophenyl)- 3-hydroxy-2- naphthalen-1-ylmethyl-2,3- dihydroisoindol- 1-one

96   NCL-00016047 2-(3-bromobenzyl)- 3-(4-chlorophenyl)- 3-hydroxy-2,3-dihydroisoindol-1-one

NCL-00016106 3-(4-chlorophenyl)- 3-(1-hydroxymethyl-cyclopropylmethoxy)- 2-naphthalen-1- ylmethyl-2,3- dihydroisoindol-1-one

49   NCL-00016107 2-(3-bromobenzyl)-3- (4-chlorophenyl)-3-(1-hydroxymethyl- cyclopropylmethoxy)- 2,3-dihydroisoindol- 1-one

47   NCL-00016655 3-hydroxy-2-(4- nitrobenzyl)-3- phenyl-2,3-dihydroisoindol-1- one

87   NCL-00016656 3-(1-hydroxymethyl- cyclopropylmethoxy)-2-(4-nitrobenzyl)-3- phenyl-2,3- dihydroisoindol-1- one

8.3  NCL-00016657 3-hydroxy-2- (4-nitrobenzyl)-3- (4-fluorophenyl)-2,3-dihydroisoindol-1-one

9.46  NCL-00016149 succinic acid mono-{1- [7-chloro-1-(4-chlorophenyl)-2-(4- nitrobenzyl)-3-oxo- 2,3-dihydro- 1H-isoindol-1-yloxymethyl] cyclopropylmethyl} ester

0.019 ± 0.009 NCL-00016659 succinic acid mono-{1- [7-chloro-1-(4-chlorophenyl)-2-(4- cyanobenzyl)-3-oxo- 2,3-dihydro- 1H-isoindol-1-yloxymethyl] cyclopropylmethyl} ester

0.102 (n = 2) NCL-00016653 succinic acid mono- {1-[2-(4-bromobenzyl)-7-chloro- 1-(4-chlorophenyl)- 3-oxo-2,3-dihydro-1H-isoindol-1- yloxymethyl] cyclopropylmethyl} ester

0.102 (n = 2) NCL-00016865 3-(4-chlorophenyl)-3- ((1-(hydroxymethyl)cyclopropyl)methoxy)- 2-(4-methylbenzyl) isoindolin-1-one

2.3  NCL-00016866 3-(4-chlorophenyl)-3- ((1-(hydroxymethyl)cyclopropyl)methoxy)- 2-(4-methoxybenzyl) isoindolin-1-one

2.8  NCL-00016895 3-(4-chlorophenyl)- 3-(2-(hydroxymethyl)allyloxy)-2-(4- nitrobenzyl)isoindolin- 1-one

0.68  NCL-00016896 3-(4-fluorophenyl)-3- ((1-(hydroxymethyl)cyclopropyl)methoxy)- 2-(4-nitrobenzyl) isoindolin-1-one

2.7  NCL-00016897 4-chloro-3-(4- chlorophenyl)-5- fluoro-3-hydroxy-2-(4-nitrobenzyl) isoindolin-1-one

34  

Of the compounds in Table 5 above, those wherein R² is hydrogen and allof R⁴-R⁷ are H are used as intermediate compounds in the preparation ofthe compounds of the invention.

Studies of the p53 binding pocket on the MDM2 protein guided the natureof the molecules synthesised. Thus the present invention provides smallmolecule inhibitors of MDM2-p53 interaction based on an isoindolinonescaffold. Preliminary screening studies, using an in vitro MDM2-p53binding assay identified the particularly useful isoindolin-1-onecompounds (Tables 2-4) as inhibitors of MDM2-p53 interaction having anIC₅₀ in the range 100-1000 nM (IC₅₀ is the concentration of a particularcompound required to inhibit 50% of a specific measured activity, inthis case inhibition of the MDM2-p53 interaction). The isoindolin-1-oneswere found to be active in the inhibition of the MDM2-p53 interaction.

The inhibitory efficacies of the compounds of the present invention havebeen determined using the ELISA assay which for the avoidance of doubtis described below.

As referred to herein “cancer” or “tumour” includes, but is not limitedto, lung cancer, small cell lung cancer, non-small cell lung cancer,gastrointestinal cancer, stomach cancer, bowel cancer, colon cancer,rectal cancer, colorectal cancer, thyroid cancer, breast cancer, ovariancancer, endometrial cancer, prostate cancer, testicular cancer, livercancer, kidney cancer, renal cell carcinoma, bladder cancer, pancreaticcancer, brain cancer, glioma, sarcoma, osteosarcoma, bone cancer, skincancer, squamous cancer, Kaposi's sarcoma, melanoma, malignant melanoma,lymphoma, or leukemia. Compounds of the present invention have beenshown to inhibit the interaction of p53 with MDM2. Such inhibition leadsto cell arrest and apoptosis.

Accordingly, the compounds of the present invention are of particularinterest for the treatment of a range of selected cancer tumours, andthe invention further provides a method for the treatment of a patientsuffering from cancer. Thus, a therapeutically effective non-toxicamount of a compound of the first and second aspects of the invention,may be suitably administered orally, parenterally (includingsubcutaneously, intramuscularly, and intravenously or topically). Theadministration will generally be carried out repetitively at intervals,for example once or several times a day.

The amount of the compound, which is required in order to be effectiveas an anti tumour agent for treating mammals will of course vary and isultimately at the discretion of the medical or veterinary practitionertreating the mammal in each particular case. The factors to beconsidered by such a practitioner include the route of administrationand pharmaceutical formulation; the mammal's body weight, surface area,age and general condition; and the chemical form of the compound to beadministered. However, a suitable effective anti tumour dose may be inthe range of about 1.0 to about 75 mg/kg bodyweight, typically in therange of about 5 to 40 mg/kg with most suitable doses being for examplein the range of 10 to 30 mg/kg. In daily treatment for example, thetotal daily dose may be given as a single dose, multiple doses, such astwo to six times per day, or by intravenous infusion for any selectedduration. For example, in the case of a 75 kg mammal, the dose rangecould be about 75 to 500 mg per day and it is expected that a typicaldose would commonly be about 100 mg per day. If discrete multiple dosesare indicated, treatment might typically be 50 mg of the compound offormula given 4 times per day in the form of a tablet capsule, liquid(for example, syrup) or injection.

The dosing will depend upon the subject or patient which may be achordate, a vertebrate, a mammal, a placental mammal, a marsupial, amonotreme (for instance, duckbilled platypus), a rodent, murine (forinstance, a mouse), a lagomorph (for instance, a rabbit), avian, canine,feline, equine, porcine, ovine (for instance, a sheep), bovine, aprimate, simian (for instance, a monkey or ape), a monkey (for instance,marmoset, baboon), an ape (for instance, gorilla, chimpanzee,orangutang, gibbon), or a human. Furthermore, the subject/patient may beany of its forms of development, for example, a foetus. Typically, thesubject is a human.

While it may be possible for the compounds of the first or secondaspects of the invention to be administered alone as the raw chemical,it is preferable to present the compound in a pharmaceuticalcomposition. Thus, the invention also provides pharmaceuticalcompositions comprising an effective amount of a compound ashereinbefore defined which forms the active therapeutic ingredient. Suchpharmaceutical compositions for medical use will be formulated inaccordance with any of the methods well known in the art of pharmacy foradministration in any convenient manner. The compounds will usually beadmixed with at least one other ingredient providing a compatiblepharmaceutically acceptable additive carrier, diluent or excipient, andmay be presented in unit dosage form.

The carrier(s) must be pharmaceutically acceptable in the sense of beingcompatible with the other ingredients of the formulation and notdeleterious to the recipient thereof.

The possible formulations include those suitable for oral, rectal,topical and parenteral (including subcutaneous intramuscular andintravenous) administration or for administration to the lung or otherabsorptive site such as the nasal passages.

All methods of formulation in making up such pharmaceutical compositionswill generally include the step of bringing a compound as defined in thefirst to third aspects of the invention into association with a carrierwhich constitutes one or more accessory ingredients. Usually, theformulations are prepared by uniformly and intimately bringing thecompound into association with a liquid carrier or with a finely dividedsolid carrier or with both and then, if necessary, shaping the productinto desired formulations.

Formulations of the present invention suitable for oral administrationmay be presented as discrete units such as capsules, cachets, tables orlozenges, each containing a predetermined amount of the compound; as apowder or granules; or a suspension in an aqueous liquid or non-aqueousliquid such as a syrup, an elixir, an emulsion or a draught. Thecompound may also be presented as bolus, electuary or paste.

A tablet may be made by compression or moulding, optionally with one ormore accessory ingredients. Compressed tablets may be prepared bycompressing, in a suitable machine, the compound in a free-flowing formsuch as a powder or granules, optionally mixed with a binder, lubricant,inert diluent, surface active or dispersing agent. Moulded tablets maybe made by moulding in a suitable machine, a mixture of the powderedcompound with any suitable carrier.

A syrup may be made by adding the compound to a concentrated, aqueoussolution of a sugar, for example sucrose, to which may be added anydesired accessory ingredients. Such accessory ingredient(s) may includeflavourings, one or more agents to retard crystallisation of the sugaror one or more agents to increase the solubility of any otheringredient, such as a polyhydric alcohol, for example glycerol orsorbitol.

Formulations for rectal administration may be presented as a suppositorywith a carrier, for instance cocoa butter.

Formulations suitable for parental administration conveniently comprisea sterile aqueous preparation of a compound of the first or secondaspects of the invention, which is typically isotonic with the blood ofthe recipient.

In addition to the aforementioned ingredients, formulations of thisinvention, for example ointments, creams and such like, may include oneor more accessory ingredients, for example a diluent, buffer, flavouringagent, binder, surface active agent, thickener, lubricant and/or apreservative (including an antioxidant) or other pharmaceutically inertexcipient.

The compounds of the present invention may also be made up foradministration in liposomal formulations, which can be prepared bymethods well-known in the art.

Where the compound is provided as part of a kit, it is typical that thekit contains a compound as defined in the first or second aspect of theinvention, or a composition comprising a compound as described herein(typically provided in a suitable container and/or with suitablepackaging); and instructions for use (for example, written instructionson how to administer the active compound or composition). Theinstructions may also include a list of indications for which the activeingredient is a suitable treatment.

The isoindolinone compounds of the present invention may be administeredalone or as a combination therapy. For instance, the compounds describedherein may also be used in combination with one another or inconjunction with other agents, for example, cytotoxic agents oranticancer agents. Examples of treatments and therapies include, but arenot limited to, chemotherapy (the administration of active agents,including, for instance, drugs, antibodies (as in immunotherapy),prodrugs (as in photodynamic therapy, GDEPT, ADEPT, etc.)); surgery;radiation therapy; photodynamic therapy; gene therapy; and controlleddiets. The particular combination would be at the discretion of thephysician who would select dosages using his common general knowledgeand dosing regimens known to a skilled practitioner.

Analytical Techniques ELISA Assay

Streptavidin-coated 96-well plates are used to immobilise abiotin-tagged IP3 p53-derived peptide (MPRFMDYWEGLN). This is a peptideanalogue derived from the p53 binding site for MDM2 (QETFSDLWKLLP). IP3has a higher affinity for MDM2 than the native peptide and has been usedelsewhere to identify antagonists of the binding between MDM2 and p53(Stoll et al 2001). Aliquots of MDM2 generated by in vitro translationare pre-incubated for 20 minutes at room temperature (i.e. 20-25° C.)with test compounds and controls, before transfer into the IP3-coated96-well plates. Following a further incubation period of 90 minutes at4° C., the plates are washed to remove unbound MDM2 and the residualbound MDM2 is detected using a primary monoclonal antibody (MDM2 Ab-1,clone IF2, Oncogene Research Products) and HRP-conjugated secondaryantibody (Goat anti-mouse, Dako PO447). The HRP (horseradish peroxidase)is measured by a chemiluminescence reaction using standard reagents(Amersham Pharmacia™ RPN 2106) and an automatic injection 96-well plateilluminometer (EG & G Berthold Microplate LB 96V).

For validation and subsequently as positive controls, IP3 & AP peptidesare used, together with the isoindolin-1-one lead compound that at thetime shows the highest degree of antagonistic activity.3-(4-Chloro-phenyl)-3-(4-hydroxy-3,5-dimethoxy-benzyloxy)-2-propyl-2,3-dihydro-isoindolin-1-one(NU8231) is currently included as a standard “lead compound” positivecontrol. AP is an octomer synthetic peptide that inhibits the p53-MDM2interaction with high potency (IC₅₀=5.0 nM) and has been reported tostimulate p53 and downstream apoptotic pathways in intact tumour celllines (Chene et al 2000). The AP peptide is included as a positivecontrol for biological evaluation of the isoindolinones in the cell freebinding assays.

All compounds are dissolved in DMSO and tested at a range ofconcentrations in the presence of a fixed final concentration of 5%DMSO. The percentage inhibition of complex formation is expressedrelative to a DMSO only control and an IC₅₀, defined as theconcentration required for 50% inhibition of MDM2-p53 complex formation,determined by interpolation.

The ELISA assay showed a standard error for n=3 independent IC₅₀determinations of 10-15% of the mean value. Thus, the variation in theIC₅₀ determination for an individual compound was much smaller than therange of values for the compounds evaluated thus far is (26.7>500 μM).

Western Blot Method

Osteosarcoma cell line SJSA-1 was plated out in 55 mm dishes at adensity of 2.5×10⁵ cells in 3 mL of RPMI 1640 medium (Sigma)supplemented with 10% foetal bovine serum (FBS, Gibco), 1% (v/v) HEPES(Gibco), 1% (v/v) sodium pyruvate (Gibco) and 1.25 g/500 ml glucose(Sigma) for 48 hours in a 37° C. humidified incubator (Sanyo, MCO 20AIC)at a CO₂ concentration of 5%.

The dishes were treated with3-(4-Chloro-phenyl)-3-(4-hydroxy-3,5-dimethoxy-benzyloxy)-2-propyl-2,3-dihydro-isoindolin-1-one(NU8231) at a final concentration of 5, 10, and 20 μM (at 1% DMSO)together with a 1% DMSO and an untreated control sample for 6 hours. Themedium was then aspirated and the dishes were washed with 3 mL of coldPBS. The cells were then lysed in 40 μL of Sodium Dodecyl Sulphate (SDS,Sigma) lysis buffer, boiled at 100° C. for 10 minutes before sonicationfor 3×5 seconds at 20 microns (Soniprep 150, MSE).

The protein concentration for each of the samples was then determinedusing BCA Protein Assay Kit (Pierce), and 1:1 loading buffer consistingof β-mercaptoethanol (Sigma) and 0.5% bromophonol-blue (Sigma) wereadded to 40 μg of protein and made up to a final volume of 30 μL andboiled for 5 minutes at 100° C.

The samples were then loaded onto a precast 4-20% gradientpolyacrylamide Tris-Glycine gels (15 wells, 1.5 mm thickness, InvitrogenLife Technologies), along with a pre-stained marker protein (SeeBlue,Invitrogen). The Gels were processed in Novex XCell (Invitrogen) at 180Vand blotted onto a High Bond C membrane (Amersham Life Science)overnight at 30V.

The membrane was then blocked for one hour at room temperature inTBS-Tween containing 5% non-fat milk (TBST-M) followed by incubationwith primary antibodies for MDM2 (MDM2-Ab1, 1:500, Oncogene), p53(p53-D07, 1:1000, Novacastra), p21 (p21 Ab1, 1:100, Oncogene) and Actin(Actin AC40, 1:1000, Sigma) in PBST-M for 1 hour.

The membrane was then washed three times in TBST (15 minutes per wash)and then incubated for an additional 1 hour with a anti mouse or arabbit horseradish peroxidase (HRP) secondary antibody (Dako, 1:1000) inPBST-M followed by a final wash consisting of six washes with TBST at 5minutes per wash. Enhanced chemiluminescence (ECL, Amersham) detectionreagents were then added onto the membrane which was exposed to a bluelight sensitive X-ray film (Fuji Photo Film Co Ltd) and developed in anautomated X-ray film processor, (Mediphot 937).

RESULTS

The invention will now be described, by way of example only, byreference to the accompanying figures, of which:

FIG. 1 is a Western Blot assay illustrating the effects of NU8293,NU8295, NU8296 and NU8297 in the SJSA-1 cell line;

FIG. 2 is a Western Blot assay illustrating the effects of NU8352,NU8353 and NU8354 compared with the effects with Nutlin-3 in the SJSA-1cell line;

FIG. 3 is a graph illustrating the effect of the NU8293, NU8296 andNU8297 isoindolinones on p53-dependent transcriptional activity measuredby a Luciferase based reporter genes assay;

FIG. 4 is a western blot illustrating the dose response effects ofNU8296 and Nutlin-3 (and NU8291, which is not part of this invention) onPARP and Caspase 3 cleavage in the SJSA-1 cell line;

FIG. 5 is a graph illustrating the growth inhibitory effects of Nutlin-3in p53 wild-type and p53 deleted versions of the HCT116 cell line withconcentrations for 50% growth inhibition shown;

FIG. 6 is a graph illustrating the effect of NU8354 as a growthinhibitor in wild-type and p53 mutant versions of the HCT116 cell linewith concentrations for 50% growth inhibition shown;

FIG. 7 is a graph illustrating the induction of Caspase 3 and 7enzymatic activity by NU8354 in the SJSA-1 cell line;

FIG. 8 is a graph illustrating the growth inhibition dose response toexposure of Nutlin-3 and NU8354 in the SJSA-1 cell line;

FIG. 9 is a graph illustrating the growth inhibition dose response toexposure of Nutlin-3 and NU8354 in the LS cell line; and

FIG. 10 is a graph illustrating the growth inhibition dose response todrug exposure of Nutlin-3 and NU8354 in the JAR cell line.

FIG. 11 is an analytical chromatogram of NU8354 on a Chiracel AD column(4.6 mm×25 cm) with 40% EtOH, pentane as eluant in isocratic mode.

FIG. 12 shows the cellular activity of Nutlin-3, NU8354 and enantiomersNU8354A and NU8354B in SJSA-1 cells by Western blotting.

NU8296 corresponds to 3-(4-chlorophenyl)-3-((1S,3R)-3-hydroxycyclopentyloxy)-2-(4-nitrobenzyl)isoindolin-1-one, a further compound inthe isoindolin-1-one series, the structure of which is given below.

Nutlin-3 is the proprietary name for(±)-4-[4,5-bis-(4-Chlorophenyl)-2-(2-isopropoxy-4-methoxyphenyl)-4,5-dihydroimidazole-1-carbonyl]-piperazin-2-one,the structure of which is given below. Nutlin-3 has been found to havean IC₅₀ of 45±4 nM and is included in the tests to provide a comparisonof the efficacy of the inventive compounds with a known inhibitor of theMDM2-p53 interaction.

FIG. 1 shows the induction of increased levels of p53, p21 and MDM2protein by treatment of MDM2 amplified SJSA-1 cells with NU8293, NU8295and NU8291 in the 1-20 μM concentration range. This is consistent withthe inhibition of MDM2-p53 binding and release of p53 activity fromnegative regulation by MDM2 in these cells, resulting in the increasedexpression and accumulation of MDM2 and p21 proteins. NU8297 did notshow any evidence of activity in this experiment.

FIG. 2 shows that NU8534 treatment results in strong induction of MDM2,p53, and p21, when compared with the positive control Nutlin-3; this isclear, for instance, from a comparison of the effects at a concentrationof 10 μM. These results are consistent with the transcriptionalactivation of p53 resulting from its release from MDM2 inhibition.

FIG. 3 shows examples of activation of p53 dependent transcriptionalactivity by NU8293, NU8296 and NU8297 measured by a luciferase basedp53-dependent reporter gene assay. This provides further evidence thatthis series of compounds specifically induces p53-dependenttranscriptional activity, consistent with the release of active p53 byinhibition of MDM2-p53 binding in intact cells. The time course ofactivation is similar to that seen with nutlin-3 and concentrationranges required are comparable.

FIG. 4 shows the dose dependence of PARP and caspase-3 cleavage inSJSA-1 cells detected after 48 hours of treatment with NU8296 incomparison to nutlin-3. The levels of PARP and caspase-3 cleavage arecomparable to those observed with nutlin-3 in the same dose range.

FIGS. 5 and 6 show that NU8534 is growth inhibitory in wild-type p53HCT116 cells, as is Nutlin-3. These effects are consistent with thetranscriptional activation of p53 and consequent induction of thep21^(WAF1) cyclin dependent kinase inhibitor and hence growth arrest.Furthermore, NU8354 displays greater growth inhibition in the p53 wtHCT116 cell line than the p53−/− HCT116 cells, as does Nutlin-3. Theseeffects demonstrate a predominantly p53-dependent mechanism of growthinhibitory activity. In addition, NU8354 shows greatest growthinhibitory activity in MDM2 amplified and p53 wt cell lines.

FIG. 7 shows that NU8354 induces Caspase 3 and 7 enzymatic activity overa 24 and 48 hr exposure in SJSA cells; this is an indication of theinduction of apoptosis and is consistent with the western blot evidenceof PARP and caspase-3 cleavage.

FIGS. 8 to 10 show growth inhibition by NU8354 in the 5-20 μMconcentration range for a panel of MDM2 amplified cell lines, which aregenerally found to be more sensitive than cell lines not amplified forMDM2.

FIG. 11 shows the analytical chromatogram of NU8354 on a Chiracel ADcolumn (4.6 mm×25 cm) with 40% EtOH, pentane as eluant in isocraticmode. The (+)-enantiomer NU8354A has a retention time of 9.8 minutes,whereas the (−)-enantiomer NU8354B elutes at 12.4 minutes. The absoluteconfiguration of the enantiomers has not been determined

FIG. 12 shows the cellular activity of Nutlin-3, NU8354 and enantiomersNU8354A and NU8354B in SJSA-1 cells by Western blotting. Nutlin-3 showsa strong dose dependent increase in MDM2, p53 and p21 levels from 1 to20 μM. A similar but weaker effect is observed for NU8354 with a maximaleffect observed at 10 μM. The activity of NU8354A is slightly weakerthan Nutlin at the 20 μM dose and significantly weaker at the lowerdoses. The NU8354B enantiomer displays little cellular activity withweak induction of p21 and MDM2 at the 20 μM dose. These results areconsistent with NU8354A being the enantiomer which confers the majorityof the biological activity of the racemate and the observed IC₅₀s in thein vitro ELISA assay.

In summary, NU8534 shows a range of cellular effects consistent with thedisruption of MDM2-p53 binding, the proposed mechanism of action. Incomparison with the positive control Nutlin-3, the effects are similaracross a panel of cell lines with differing p53 and MDM2 status, interms of p53 activation, growth inhibition and apoptosis.

Synthetic Data

The present invention will now be described further by way of exampleonly. The following examples and description of stages in syntheticroutes of preparation of various compounds of interest serve further toillustrate the present invention.

3-(4-Fluorophenyl)-3-hydroxy-2-propyl-2,3-dihydroisoindol-1-one

THF (25 mL) was added to 2-(4-fluorobenzoyl)benzoic acid (5 g, 20.4mmol) followed by thionyl chloride (2.97 mL, 40.9 mmol) and a catalyticamount of DMF (3 drops). The system was stirred under nitrogen for 4 hat room temperature. Removal of the solvent gave3-chloro-3-(4-fluorophenyl)-3H-isobenzofuran-1-one as a colourless oil(5.35 g, 20.4 mmol, 100%).

Distilled THF (25 mL) was added to3-chloro-3-(4-chlorophenyl)-3H-isobenzofuran-1-one (5.35 g, 20.4 mmol)followed by n-propylamine (1.85 mL, 22.5 mmol), triethylamine (2.85 mL,26.5 mmol) resulting in the formation a creamy white/yellow precipitate.The mixture was stirred at room temperature under nitrogen for 4 h thenthe solvent was removed under vacuum. The residue was taken up in ethylacetate (30 mL), washed with water (3×20 mL), brine (10 mL), dried(MgSO₄) and evaporated. Recrystallisation (ethyl acetate) gave the titlecompound as a white solid (4.35 g, 15.2 mmol, 75%); R_(f)=0.48 (40:60:EtOAc: petrol). mp 172.3-174.6° C. λ_(max) (CH₃OH)/nm 210. IR: 3231,2965, 1673, 1602, 1504, 1407, 1223 cm⁻¹. ¹H NMR: (300 MHz, d₆-DMSO)δ0.75 (t, 3H, J=7.4 Hz, CH₂—CH₂—CH₃), 1.42 (m, 2H, N—CH₂—CH₂), 2.87 (m,1H, N—CH₂), 3.14 (m, 1H, N—CH₂), 7.15 (m, 2H, Ar—H), 7.25 (m, 1H, Ar—H),7.35 (m, 2H, Ar—H), 7.53 (dquin, 2H, J=7.4, 1.4 Hz, Ar—H), 7.71 (m, 1H,Ar—H). ¹³C NMR: (75 MHz, d₆-DMSO) g 11.8, 22, 90.4, 115.4, 115.7, 122.7,123, 128.3, 128.4, 129.5, 130.8, 132.7, 136.8, 136.9, 149.7, 160.5,162.2, 163.7, 166.8. LCMS (ESI+) m/z=161.1, 227.1, 268.1, 286.1 [M+H]⁺.Anal. Calcd. for C₁₇H₁₆FNO₂: C, 71.56; H, 5.65; N, 4.91%. Found C,71.61; H, 5.70; N, 4.99%. HRMS (EI) m/z Calcd. for C₁₇H₁₆FNO₂: 285.1165.Found 285.1166.

3-(4-Fluorophenyl)-3-hydroxycyclopentyloxy)-2-propyl-2,3-dihydroisoindol-1-one

Distilled THF (20 mL) was added to3-(4-fluorophenyl)-3-hydroxy-2-propyl-2,3-dihydroisoindol-1-one (200 mg,0.7 mmol) followed by thionyl chloride (0.06 mL, 0.84 mmol) and acatalytic amount of DMF (3 drops). The mixture was stirred at roomtemperature under nitrogen for 4 h and monitored by TLC. Removal of thesolvent under vacuum gave3-chloro-3-(4-fluorophenyl)-2-propyl-2,3-dihydroisoindol-1-one as acolourless oil (212 mg, 0.69 mmol, 100%) which was used immediatelywithout further purification.

Distilled THF was added to3-chloro-3-(4-fluorophenyl)-2-propyl-2,3-dihydroisoindol-1-one (212 mg,0.69 mmol) followed by 1,3-cyclopentanediol (0.65 mL, 6.9 mmol). Themixture was stirred at room temperature under nitrogen for 4 h andmonitored by TLC. On completion the solvent was removed under vacuum,the residue was taken up in ethyl acetate (30 mL), washed with water(3×20 mL), brine (10 mL) and dried (MgSO₄). The solvent was removed togive the crude product. HPLC(H₂O:MeOH, 270 nm) gave NU8279 as a clearglass (126 mg, 0.34 mmol, 49%); R_(f)=0.21 (40:60: EtOAc:petrol).λ_(max) (CH₃OH)/nm 220.5. IR: 3387, 2936, 1683, 1604, 1505, 1366 cm⁻¹.¹H NMR: (300 MHz, d₄-MeOH) 80.77 (t, 3H, J=7.4 Hz, CH₂—CH₂—CH₃), 1.15(m, 1H, N—CH₂—CH₂), 1.32 (m, 1H, N—CH₂—CH₂), 1.40-2.05 (m, 6H,cyclopentane), 3.12 (m, 1H, N—CH₂), 3.29 (m, 1H, N—CH₂), 3.90 (m, 1H,cyclopentane), 4.31 (m, 1H, cyclopentane), 7.07 (t, 2H, J=9 Hz, Ar—H),7.23 (m, 1H, Ar—H), 7.39 (m, 2H, Ar—H), 7.60 (m, 2H, Ar—H), 7.87 (m, 1H,Ar—H). ¹³C NMR: (125 MHz, d₄-MeOH) 812.2, 22.9, 32.7, 33.1, 34.2, 43.1,44.3, 44.8, 72.8, 73, 75.7, 96.5, 116.3, 116.6, 124.3, 125.7, 130,130.1, 131.6, 133.6, 134.1, 137.1, 148.1, 166.2, 170.7. LCMS (ESI+)m/z=227.1, 268.1, 370.3 [M+H]⁺, 392.3 [M+Na]⁺. HRMS (EI) m/z Calcd. forC₂₂H₂₄FNO₃: 369.1740. Found 369.1737.

3-(4-Chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one

THF (25 mL) was added to 2-(4-chlorobenzoyl)benzoic acid (1 g, 3.8 mmol)followed by thionyl chloride (0.55 mL, 7.6 mmol) and a catalytic amountof DMF (3 drops). The system was stirred under nitrogen for 4 h at roomtemperature and monitored by TLC. Removal of the solvent gave3-chloro-3-(4-chlorophenyl)-3H-isobenzofuran-1-one as a colourless oil(1.06 g, 3.8 mmol, 100%).

Distilled THF (25 mL) was added to3-chloro-3-(4-chlorophenyl)-3H-isobenzofuran-1-one (3.2 g, 11.5 mmol),4-nitrobenzylamine hydrochloride (2.3 g, 12.6 mmol), and triethylamine(4.8 mL, 34.5 mmol). The mixture was stirred at room temperature undernitrogen for 4 h and monitored by TLC. On completion the solvent wasremoved under vacuum, the residue was taken up in ethyl acetate (30 mL),washed with water (3×20 mL), brine (10 mL) and dried (MgSO₄). Thesolvent was removed under vacuum. Recrystallisation (ethyl acetate) gave3-(4-Chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-oneas a light yellow solid (2.95 g, 7.47 mmol, 65%); R_(f)=0.4 (40:60:EtOAc:petrol). mp 197.1-199.7° C. λ_(max) (CH₃OH)/nm 225. IR: 3215,1676, 1517, 1395, 1341 cm⁻¹. ¹H NMR: (300 MHz, d₆-DMSO) δ 4.35 (d, 1H,J=16.3 Hz, N—CH₂), 4.61 (d, 1H, J=16.3 Hz, N—CH₂), 7.28 (m, 4H, Ar—H),7.45 (m, 3H, Ar—H), 7.58 (m, 2H, Ar—H), 7.79 (m, 1H, Ar—H), 8.05 (m, 2H,Ar—H). ¹³C NMR: (75 MHz, d₆-DMSO) δ 42.1, 90.5, 123.1, 123.3, 128.4,128.7, 129.1, 129.9, 130.3, 133.2, 133.3, 138.9, 146.4, 146.5, 149.4,167.1. LCMS (ESI+) m/z=307.2, 368.2, 377.1. Anal. Calcd. forC₂H₁₅ClN₂O₄: C, 63.89; H, 3.83; N, 7.10%. Found C, 63.78; H, 3.92; N,7.12%. HRMS (EI) m/z Calcd. for C₂₁H₁₅ClN₂O₄: 394.0720. Found 394.0714.

3-(4-Chlorophenyl)-2-[1-(4-chlorophenyl)-ethyl]-3-hydroxy-2,3-dihydroisoindol-1-one

To a solution of 2-(4-chlorobenzoyl)benzoic acid (5 g, 19.2 mmol, 1equiv.) in dry THF (20 mL) was added under nitrogen atmospherethionylchloride (3.0 mL, 38.3 mmol, 2 equiv.) and 3 drops of anhydrousDMF. The reaction mixture was stirred for 4 h at room temperature andconcentrated in vacuo. The resulting pale yellow oil was taken up in dryTHF (20 mL), and (S)-4-chloro-α-methylbenzylamine (2.43 g, 15.6 mmol,1.1 equiv.) and DIPEA (3.49 mL, 21.1 mmol, 1.1 equiv.) were added undernitrogen atmosphere. The reaction mixture was stirred overnight at roomtemperature and the solvents were removed in vacuo. The residue wastaken up in EtOAc (100 mL), filtered, and the filtrate washed with water(3×50 mL) and brine (1×50 mL). The organic layer was dried over MgSO₄,filtered and concentrated in vacuo to afford a solid, which wasrecrystallised (EtOAc/Petrol) as a white crystalline powder (3.10 g,55%). ¹H NMR (300 MHz, CDCl₃): Mixture of two diastereoisomers:7.74-7.09 (m, 13H, Ar—H and OH), 4.52 and 4.45 (q, J=6.9 Hz, 1H, CH*),1.75 and 1.48 (d, J=7.2 Hz, 3H, CH₃). ¹³C NMR (75 MHz, CDCl₃): Mixtureof two diastereoisomers: 166.94, 166.43, 149.17, 149.11, 142.01, 141.51,139.65, 139.33, 133.24, 133.14, 132.90, 131.76, 131.60, 131.54, 131.47,129.79, 129.71, 129.52, 128.67, 128.62, 128.26, 127.88, 127.84, 123.13,122.74, 122.70, 91.19, 90.94, 51.22, 50.46, 20.21, 18.24. FTIR: 3103(OH), 1665 (C═O) cm⁻¹. m/z (ES): 398 [M+H]⁺. Anal.: calc. forC₂₂H₁₇Cl₂NO₂+0.3H₂O: C, 65.45; H, 4.40; N, 3.47. Found: C, 65.00; H,4.30; N, 3.60.

General Procedure A

To a solution of the corresponding isoindolone (1.37 mmol, 1 equiv.) indry THF (10 mL) was added under nitrogen atmosphere, thionylchloride(214 μL, 2.75 mmol, 2 equiv.) and 3 drops of DMF. The reaction mixturewas stirred for 4 h at room temperature and concentrated in vacuo. Theresulting pale yellow oil was taken up in dry THF (10 mL), and thealcohol (2.75 mmol, 2 equiv) and potassium carbonate (380 mg, 2.75 mmol,2 equiv.) were added. The reaction mixture was stirred overnight at roomtemperature and the solvents were removed in vacuo. The residue wastaken up in EtOAc (50 mL) and washed with water (3×25 mL) and brine(1×25 mL). The organic layer was dried over MgSO₄, filtered andconcentrated in vacuo to afford an oil which was purified by flashchromatography (silica; EtOAc/petrol).

General Procedure B: Synthesis of3-Alkoxy-3-(4-chlorophenyl)isoindolin-1-ones

To a solution of the appropriate3-chloro-3-(4-chlorophenyl)isoindolin-1-one, THF was added theappropriate alcohol (5.0 mol equiv unless stated otherwise) and K₂CO₃(5.0 mol equiv unless stated otherwise). The mixture was allowed to stirat room temperature for 4 hours under nitrogen and monitored by TLC.Upon completion the mixture was extracted with EtOAc (15 mL), washedwith saturated brine (3×10 mL), water (3×10 mL) and dried (MgSO₄).Removal of the solvent under reduced pressure yielded the crude3-alkoxy-2,3-dihydroisoindolin-1-one.

General Procedure B1: Synthesis of 3-alkoxy-2,3-dihydroisoindolin-1-ones

To a solution of cis-cyclopentane or cis-cyclopentene diol (2.5 equiv)and K₂CO₃ (2.5 equiv) in THF (3 mL) was added the appropriate3-chloro-3-(4-chlorophenyl)isoindolin-1-one (1 equiv) in THF (3 mL)dropwise over 3 hours with stirring under nitrogen at room temperature.The solution was stirred for a further hour and monitored by TLC. Uponcompletion the mixture was extracted with EtOAc (15 mL), washed withsaturated brine (3×10 mL), water (3×10 mL) and dried (MgSO₄). Thesolvent was removed under reduced pressure to yield the crude3-alkoxy-2,3-dihydroisoindolin-1-one.

3-(4-Chlorophenyl)-3-hydroxy-2-(4-chlorobenzyl)-isoindolin-1-one

Distilled THF (25 mL) was added to3-chloro-3-(4-chlorophenyl)-3H-isobenzofuran-1-one (1.071 g, 3.84 mmol)followed by triethylamine (855.1.1 mg, 8.45 mmol, 1.178 mL) andpara-chlorobenzylamine (543.36 mg, 3.84 mmol, 0.467 mL) resulting in theformation of a white precipitate. The mixture was stirred at roomtemperature under nitrogen for 4 hours and monitored by TLC. Uponcompletion the mixture was then extracted with EtOAc (15 mL) and washedwith saturated sodium bicarbonate (3×10 mL), water (3×10 mL) and dried(Na₂SO₄). The solvent was removed under reduced pressure and theresultant precipitate recrystallised (EtOAc, petrol) to give the titleproduct as a fine white crystalline solid (1.191 g, 3.20 mmol, 83%). ¹HNMR: (300 MHz, DMSO) δ ppm 4.23 (d, 1H, J=15.59, H₉) 4.45 (d, 1H,J=15.62H_(9′)) 7.25 (m, 9H, H₁—H₅, H₁₀—H₁₃) 7.56 (m, 2H, H₆—H₇) 7.76 (d,1H, H₈). ¹³C NMR: (75 Hz, DMSO); δ 42.09 (N—CH₂), 90.55 (O—C—N), 122.98,123.19, 128.02, 128.39, 128.57, 129.76, 130.21, 130.66, 131.60, 133.07,133.15, 137.44, 139.33, 149.52 (Ar), 167.16 (C═O). Mp: 156.2-156.9° C.IR: 1467, 1661, 3184 cm⁻¹.

3-(4-Chlorophenyl)-3-hydroxy-2-propylisoindolin-1-one

Distilled THF (20 mL) was added to3-chloro-3-(4-chlorophenyl)-3H-isobenzofuran-1-one (535.4 mg, 1.91 mmol)followed by triethylamine (398.9 mg, 0.549 mL, 3.94 mmol) andn-propylamine (159.8 mg, 0.22 mL, 1.792 mmol). The mixture was stirredat room temperature under nitrogen for 4 hours and monitored by TLC.Upon completion the mixture was then extracted with EtOAc (15 mL) andwashed with saturated sodium bicarbonate (3×10 mL), water (3×10 mL) anddried (Na₂SO₄). Recrysallisation from a minimum amount of boiling ethylacetate and an excess of petrol yielded the title product as a whitecrystalline solid (425 mg, 1.409 mmol, 77%). ¹H NMR: (300 Hz, CDCl₃) δppm 0.76 (t, 3H, H₁₁), 1.45 (m, 2H, H₁₀), 2.86 (m, 1H, H₉), 3.36 (m, 1H,H_(9′)), 7.16 (d, 1H, H₅), 7.38 (dd, 4H, H₁—H₄), 7.45 (m, 2H, H_(6—)H₇),7.70 (d, 1H, H₈). ¹³C NMR: (75 Hz, DMSO); 811.87 (N—CH₂—CH₂—CH₃), 21.96(N—CH₂—CH₂), 40.69 (N—CH₂), 90.50 (O—C—N), 122.75, 122.99, 128.24,128.76, 129.58, 131.04, 131.24, 132.73, 133.13, 139.94, 149.59 (Ar),166.96 (C═O). Mp: 201.5-201.7° C. IR: 1466, 1608, 1664, 2968, 3157 cm⁻¹.

2-Benzyl-3-(4-chlorophenyl)-3-hydroxy-2,3-dihydroisoindolin-1-one

Distilled THF (25 mL) was added to3-chloro-3-(4-chlorophenyl)-3H-isobenzofuran-1-one (1.042 g, 3.84 mmol)followed by triethylamine (777.1 mg, 7.68 mmol, 1.06 mL)) andbenzylamine (616 mg, 5.76 mmol, 0.79 mL) resulting in the formation of ayellow/cream precipitate. The mixture was stirred at room temperatureunder nitrogen for 4 hours and monitored by TLC. Upon completion themixture was then extracted with EtOAc (15 mL) and washed with saturatedsodium bicarbonate (3×10 mL), water (3×10 mL) and dried (Na₂SO₄).Recrystallisation of the cream precipitate from a minimum amount ofboiling ethyl acetate and excess petrol to yielded the title product asa white crystalline solid (1.0378 g, 2.96 mmol, 77% yield). ¹H NMR: (300Hz, CDCl₃) δ ppm 4.24 (d, 1H, J=15.48 Hz, H₉), 4.42 (d, 1H, J=15.48 Hz,H_(9′)), 7.16 (m, 5H, H₁₀—H₁₄), 7.25 (m, 5H, H₁—H₅), 7.56 (m, 2H,H₆—H₇), 7.75 (d, 1H, H₈). ¹³C NMR: (75 Hz, DMSO); δ 42.80 (N—CH₂), 90.60(O—C—N), 122.95, 123.18, 126.78, 128.07, 128.35, 128.41, 128.51, 129.70,130.81, 132.97, 133.07, 138.42, 139.46, 149.59 (Ar), 167.20 (C═O). IR:1463, 1664, 2936, 3285 cm⁻¹.

3-(4-Chlorophenyl)-3-hydroxy-2-(4-methylbenzyl)-isoindolin-1-one

Distilled THF (25 mL) was added to3-chloro-3-(4-chlorophenyl)-3H-isobenzofuran-1-one (1.071 g, 3.84 mmol)followed by triethylamine (855.1.1 mg, 8.45 mmol, 1.178 mL) andpara-methylbenzylamine (465.3 mg, 3.84 mmol, 0.489 mL) resulting in theformation of a bright yellow precipitate. The mixture was stirred atroom temperature under nitrogen for 4 hours and monitored by TLC. Uponcompletion the mixture was then extracted with EtOAc (15 mL) and washedwith saturated sodium bicarbonate (3×10 mL), water (3×10 mL) and dried(Na₂SO₄). Recrystallisation of the cream/yellow residue from a minimumamount of boiling ethyl acetate and excess petrol yielded the titleproduct as a fine pale yellow crystalline solid (1.090 g, 3.10 mmol,81%). ¹H NMR: (300 MHz, DMSO) δ ppm 2.21 (s, 3H, HO, 4.20 (d, 1H,J=15.62 Hz, H₉), 4.40 (d, 1H, J=15.36 Hz, H_(9′)), 6.95 (d, 2H J=7.97Hz, H₁₁, H₁₃), 7.04 (d, 2H J=8.00 Hz, H₁₀—H₁₄) 7.27 (m, 5H, H₁—H₅), 7.55(m, 2H, H₆—H₇) 7.74 (d, 1H, H₈). ¹³C NMR: (75 Hz, DMSO) δ 20.90 (CH₃),42.53 (N—CH₂), 90.57 (O—C—N), 122.89, 123.15, 128.36, 128.38, 128.61,129.66, 130.85, 132.91, 133.03, 135.37, 135.84, 139.51 (Ar), 167.10(C═O). IR: 1398, 1468, 1660, 2921, 3138 cm⁻¹.

2-(4-(Aminomethyl)benzonitrile)-3-(4-chlorophenyl)-3-hydroxyisoindolin-1-one

Distilled THF (25 mL) was added to3-chloro-3-(4-chlorophenyl)-3H-isobenzofuran-1-one (1.071 g, 3.84 mmol)followed by triethylamine (777.14 mg, 7.68 mmol, 1.07 mL) and4-(aminomethyl)benzonitrile (507.6 mg, 3.84 mmol). The mixture wasstirred at room temperature under nitrogen for 4 hours and monitored byTLC. Upon completion the mixture was then extracted with EtOAc (15 mL)and washed with saturated sodium bicarbonate (3×10 mL), water (3×10 mL)and dried (Na₂SO₄). Removal of solvent after washing produced a viscousorange oil. Trituration under petrol yielded a yellow/orange solid whichwas recrystallised from a minimum amount of boiling ethyl acetate andexcess petrol to produce the title product as a fine pale yellowcrystalline solid (665 mg, 1.83 mmol, 50%). ¹H NMR: (300 MHz, DMSO) δppm 4.31 (d, 2H, J=16.12 Hz, H₉), 4.55 (d, 2H J=16.15 Hz, H_(9′)), 7.26(m, 5H, H₁—H₅), 7.37 (d, 2H J=8.29 Hz, H₁₀, H₁₃), 7.58 (m, 2H, H₆—H₇),7.64 (d, 2H J=8.26 Hz, H₁₁—H₁₂), 7.77 (d, 1H, H₈). ¹³C NMR: (75 Hz,DMSO); δ 42.51 (N—CH₂), 60.02 (CN) 90.57 (O—C—N), 109.82, 119.08,123.06, 123.25, 128.40, 128.62, 129.16, 129.83, 130.54, 132.03, 133.19,133.26, 133.52, 139.19, 144.24, 149.48 (Ar), 167.23 (C═O). IR: 1397,1655, 2227 cm⁻¹.

4-((1-(4-chlorophenyl)-1-(4-hydroxycyclopent-2-enyloxy)-3-oxoisoindolin-2-yl)methyl)benzonitrile(NU8292)

Distilled THF (10 mL) was added to2-(4-(aminomethyl)benzonitrile)-3-(4-chlorophenyl)-3-hydroxyisoindolin-1-one(400 mg, 1.10 mmol), thionyl chloride (288.9 mg, 2.42 mmol, 0.18 mL) andcatalytic DMF (3 drops) as for general procedure B1.3-Chloro-3-(4-chlorophenyl)-2-(4-(aminomethyl)benzonitrile)isoindolin-1-one was produced as a viscous clear oil (419 mg, 1.10 mmol)which was used immediately without further purification. Distilled THF(3 mL) was added to3-chloro-3-(4-chlorophenyl)-2-(4-(aminomethyl)benzonitrile)isoindolin-1-one(419 mg, 1.10 mmol) and the resultant solution added dropwise tocis-cyclopentene diol (275 mg, 2.75 mmol) and dried K₂CO₃ (380.05 mg,2.75 mmol) in distilled THF (3 mL) as for general procedure B1. Removalof the solvent yielded the crude product as a yellow oil (364 mg).Purification by flash column chromatography yielded the title product asa yellow oily solid (105.4 mg, 0.23 mmol, 21%). ¹H NMR: (300 MHz, DMSO)δ ppm 1.31 (m, 1H, H_(15/15′))_(,) 1.73, 2.11 (m, 1H, H_(15/15′)), 3.89(br s, 1H, OH), 4.22 (m, 1H, H₁₆), 4.42 (d, 1H, J=7.10 Hz, H_(9/9′)),4.52 (d, 1H J=7.03 Hz, H_(9/9′)), 4.98, 5.04 (dd, 1H, H_(17/18)), 5.01,5.27 (dd, 1H, H_(17/18)), 5.75 (m, 1H, H₁₄), 7.24 (m, 7H, H₁—H_(5,)H_(10,) H₁₃) 7.62 (m, 4H, H₆/₇, H_(11/12)), 7.87 (d, 1H, H₈). Mp:126.4-127.8° C. IR: 1382, 1609, 2229, 3061 cm⁻¹. HR-MS (ED: Calculatedmass: [M+Na]⁺479.1133. Found: 479.1134.

3-(4-Chlorophenyl)-3-(4-hydroxycyclopent-2-enyloxy)-2-(4-chlorobenzyl)isoindolin-1-one (NU8293)

Distilled THF (10 mL) was added to3-(4-Chlorophenyl)-3-hydroxy-2-(4-chlorobenzyl)-isoindolin-1-one (200mg, 0.54 mmol), thionyl chloride (141.37 mg, 1.18 mmol, 0.09 mL) andcatalytic DMF (3 drops) as for general procedure B1.3-Chloro-3-(4-chlorophenyl)-2-(4-chlorobenzyl)isoindolin-1-one wasproduced as a viscous colourless oil (210 mg, 0.54 mmol) which was usedimmediately without further purification. Distilled THF (3 mL) was addedto 3-chloro-3-(4-chlorophenyl)-2-(4-chlorobenzyl)isoindolin-1-one (210.8mg, 0.54 mmol) and the resultant solution added dropwise tocis-cyclopentene diol (135 mg, 1.35 mmol) and dried potassium carbonate(186.6 mg, 1.35 mmol) in distilled THF (3 mL) as for general procedureB1. Removal of the solvent yielded the crude product as a yellow oil(224 mg). The sample was purified by flash column chromatography(EtOAc:Petrol, 40:60) to yield the title product as a yellow viscousoil. (103.9 mg, 0.23 mmol, 43%). ¹H NMR: (300 MHz, DMSO) δ ppm 1.27 (m,1H, H_(15/15′)), 1.41, 1.68 (m, 1H, H_(15/15′)) 3.86 (m, 1H, H₁₆), 4.16(m, 1H, H₁₄) 4.31 (d, 1H, J=15.49, H_(9/9′)) 4.50 (d, 1H,J=15.48H_(9/9)) 4.93, 5.75 (dd, 1H, H_(17/18)) 5.25, 5.70 (dd, 1H,H_(17/18)), 7.21 (m, 9H, H₁—H₅, H₁₀—H₁₃) 7.63 (m, 2H, H₆—H₇) 7.85 (d,1H, H₈). IR: 1467, 1683, 2926, 3397 cm⁻¹.

3-(4-Chlorophenyl)-3-(4-hydroxycyclopent-2-enyloxy)-2-propylisoindolin-1-one(NU8294)

Distilled THF (5 mL) was added to3-(4-chlorophenyl)-3-hydroxy-2-propylisoindolin-1-one (104 mg, 0.33mmol), thionyl chloride (86.83 mg, 0.73 mmol, 0.05 mL) and catalytic DMF(3 drops) as for general procedure B1.3-Chloro-3-(4-chlorophenyl)-2-propylisoindolin-1-one was produced as anorange oil (105 mg, 0.33 mmol) which was used immediately withoutfurther purification. Distilled THF (6 mL) was added to3-chloro-3-(4-chlorophenyl)-2-propylisoindolin-1-one (211.2, 0.66 mmol),cis-cyclopentene diol (330 mg, 3.3 mmol) and potassium carbonate (456mg, 3.3 mmol) as for general procedure B. Removal of the solvent yieldedthe crude product as a colourless oil (160.2 mg). The sample waspurified by flash column chromatography (EtOAc:Petrol, 40:60) to yieldthe title product as an colourless viscous oil. (129 mg, 0.43 mmol, 51%yield). ¹H NMR: (300 Hz, DMSO) δ ppm 0.76 (t, 3H, H₁₁), 1.36 (m, 1H,H₁₃/H_(13′))_(,) 1.49, 1.76 (m, 1H, H₁₃/H_(13′)), 3.12 (m, 2H H₁₀) 3.96(m, 1H, H₉/H_(9′)), 4.26 (m, 1H, H₉/H_(9′)), 5.29, 5.87, (dd, 1H,H₁₅/H₁₆) 5.74, 5.79 (dd, 1H, H₁₅/H₁₆) 7.23 (d, 1H,), 7.38 (m, 4H,H₁—H₅), 7.61 (m, 2H, H₆H₇), 7.76 (d, 1H, H₈). IR: 1365, 1682, 2969, 3363cm⁻¹.

3-(4-Chlorophenyl)-3-(4-hydroxycyclopent-2-enyloxy)-2-(4-methylbenzyl)isoindolin-1-one (NU8295)

Distilled THF (10 mL) was added to3-(4-chlorophenyl)-3-hydroxy-2-(4-methylbenzyl)-isoindolin-1-one (400mg, 1.13 mmol), thionyl chloride (295.8 mg, 2.48 mmol, 0.18 mL) andcatalytic DMF (3 drops) as for general procedure B1.3-Chloro-3-(4-chlorophenyl)-2-(4-methylbenzyl)isoindolin-1-one wasproduced as a viscous yellow oil (418 mg, 1.13 mmol) which was usedimmediately without further purification. Distilled THF (3 mL) was addedto 3-chloro-3-(4-chlorophenyl)-2-(4-methylbenzyl)isoindolin-1-one (418.1mg, 1.13 mmol) and the resultant solution added dropwise tocis-cyclopentene diol (285 mg, 2.85 mmol) and dried K₂CO₃ (390.41 mg,2.85 mmol) in distilled THF (3 mL) as for general procedure B1. Removalof the solvent yielded the crude product as a green oil (359 g).Purification by flash column chromatography yielded the title product asa colourless viscous oil (242.1 mg, 0.55 mmol, 50%) ¹H NMR: (300 MHz,DMSO) δ ppm 1.19 (m, 1H, H_(16/16′)), 1.67, 2.05 (m, 1H, H_(16/16′))2.21 (s, 3H, H₁₂), 3.85 (m, 1H, HO, 4.06 (d, 1H, J=15.21 Hz, H_(9/9′)),4.32 (m, 1H, H₁₅) 4.53 (d, 1H, J=15.21 Hz, H_(9/9′)), 4.81, 5.74, (dd,1H, H_(18/19)) 5.20, 5.66 (dd, 1H, H_(18/19)), 6.95 (m, 4H, H₁₀, H₁₁,H₁₃, H₁₄), 7.25 (m, 5H, H₁—H₅), 7.65 (m, 2H, H₆—H₇) 7.74 (d, 1H, H₈).IR: 1380, 1467, 1699, 2922 cm⁻¹. HR-MS (EI): Calculated mass:[M+H]⁺446.1517. Found: 446.1517.

3-(4-Chlorophenyl)-3-(4-hydroxycyclopent-2-enyloxy)-2-(4-nitrobenzyl)isoindolin-1-one (NU8297)

Distilled THF (10 mL) was added to3-(4-chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl)isoindolin-1-one (200 mg,0.52 mmol), thionyl chloride (136.8 mg, 1.15 mmol, 0.08 mL) andcatalytic DMF (3 drops) as for general procedure B1.3-Chloro-3-(4-chlorophenyl)-2-(4-nitrobenzyl)isoindolin-1-one wasproduced as a viscous yellow oil (208.5 mg, 0.52 mmol) which was usedimmediately without further purification. Distilled THF (3 mL) was addedto 3-chloro-3-(4-chlorophenyl)-2-(4-nitrobenzyl)isoindolin-1-one (208.5mg, 0.52 mmol) and the resultant solution added dropwise tocis-cyclopentene diol (260 mg, 2.6 mmol) and dried potassium carbonate(359 mg, 2.6 mmol) in distilled THF (3 mL) as for general procedure B1.Removal of the solvent yielded the crude product as a yellow oil (262mg). The sample was purified by flash column chromatography(EtOAc:Petrol, 40:60) to yield the title product as a yellow viscousoil. (211 mg, 44.4 mmol, 85%). ¹H NMR: (300 MHz, DMSO) δ ppm 1.34, (m,1H, H₁₅H₁₆) 1.74, 2.2 (m, 1H, H₁₅/H₁₆) 4.22 (dt, 1H, H₁₆) 4.40 (d, 1H,J=15.99 Hz, H₉/H_(9′)), 4.63 (d, 1H, J=16.02 Hz, H₉/H_(9′)), 4.93, 4.98,(dd, 1H, H₁₇/H₁₈), 5.15, 5.29 (dd, 1H, H₁₇/H₁₈), 5.76 (m, 1H, H₁₄) 7.26(m, 7H, H₁—H₅, H₁₀, H₁₃), 7.66 (m, 2H, H₆—H₇), 7.88 (d, 1H, H₈), 8.00(m, 2H₁₁H₁₂).

2-Benzyl-3-(4-chlorophenyl)-3-(4-hydroxycyclopent-2-enyloxy)isoindolin-1-one (NU8298)

Distilled THF (25 mL) was added to2-benzyl-3-(4-chlorophenyl)-3-hydroxy-2,3-dihydroisoindolin-1-one (400mg, 1.145 mmol), thionyl chloride (299.6 mg, 2.51 mmol, 0.18 mL) andcatalytic DMF (3 drops) as for general procedure B1.3-Chloro-3-(4-chlorophenyl)-2-benzylisoindolin-1-one was produced as acolourless oil (421 mg, 1.145 mmol) which was used immediately withoutfurther purification. Distilled THF (25 mL) was added to3-chloro-3-(4-chlorophenyl)-2-benzylisoindolin-1-one (421 mg, 1.145mmol), cis-cyclopentene diol (572 mg, 5.725 mmol) and potassiumcarbonate (792.9 mg, 5.725 mmol) as for general procedure B. Removal ofthe solvent yielded the crude product as a pink oil (354 mg). The samplewas purified by flash column chromatography (EtOAc:Petrol, 40:60) toyield the title product as a cream oily solid (277.5 mg, 0.643 mmol, 56%yield). ¹H NMR: (300 Hz, CDCl₃) δ ppm 0.84, 1.66 (m, 1H, H₁₆/H_(16′))1.26 (m, 1H, H₁₆/H_(16′)) 3.85 (m, 1H, H₁₇) 3.36 (m, 1H, H₁₅) 4.15 (d,1H, J=15.27 Hz, H₉/H_(9′)), 4.62 (d, 1H, J=15.31 Hz, H₉/H_(9′)), 4.68,5.73 (dd, 1H, H₁₈/H₁₉), 5.21, 5.64 (dd, 1H, H₁₈/H_(l9)), 7.18 (m, 10H,H₁—H₅, H₁₀—H₁₄), 7.62 (m, 2H, H₆—H₇), 7.83 (d, 1H, H₈). Mp: 63.4-63.9°C. IR: 1489, 1683, 3061, 3379

3-(4-Chlorophenyl)-3-(4-hydroxybut-2-enyloxy)-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(NU8350)

The named compound was synthesised from3-(4-Chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(400 mg, 1.01 mmol, 1 equiv.) and cis-butenediol (445 mg, 5.05 mmol, 5equiv.) using General Procedure A and obtained as a yellow oil (272 mg,58%). ¹H NMR (300 MHz, CDCl₃): 8.04-8.00 (m, 2H, O₂N—C—CH), 7.93-7.91(m, 1H, C(O)═C═CH), 7.57-7.51 (m, 2H, Ar—H), 7.39-7.36 (m, 2H, Ar—H),7.23-7.12 (m, 5H, Ar—H), 5.62-5.53 (m, 1H, OCH₂CH), 5.35-5.26 (m, 1H,OCH₂CH), 4.64 and 4.26 (dd: AB, J=15.0 Hz, 2H, N—CH₂), 3.79 (d, J=6.6Hz, HO—CH₂), 3.48-3.29 (m, 2H, O—CH₂). ¹³C NMR (75 MHz, CDCl₃): 168.17,147.45, 145.07, 144.60, 136.86, 134.98, 133.07, 132.00, 131.44, 130.24,130.02, 128.71, 127.99, 126.79, 123.89, 123.35, 95.00, 59.20, 58.48,42.57. m/z (ES): 465 [M+H]⁺. Anal.: calc. for C₂₅H₂₁ClN₂O₅: C, 64.59; H,4.55; N, 6.02. Found: C, 64.39; H, 4.67; N, 5.67.

3-(4-Chlorophenyl)-3-(4-hydroxybut-2-enyloxy)-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(NU8351)

The named compound was synthesised from3-(4-Chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(400 mg, 1.01 mmol, 1 equiv.) and cis/trans-butenediol (445 mg, 5.05mmol, 5 equiv.) using General Procedure A and obtained as a yellow oil(291 mg, 62%). ¹H NMR (300 MHz, CDCl₃): 8.04-8.01 (m, 2H, O₂N—C—CH),7.94-7.91 (m, 1H, C(O)═C═CH), 7.57-7.51 (m, 2H, Ar—H), 7.40-7.36 (m, 2H,Ar—H), 7.23-7.13 (m, 5H, Ar—H), 5.62-5.53 (m, 1H, OCH₂CH), 5.35-5.27 (m,1H, OCH₂CH), 4.64 and 4.26 (dd: AB, J=15.0 Hz, 2H, N—CH₂), 3.79 (d,J=6.6 Hz, HO—CH₂), 3.48-3.29 (m, 2H, O—CH₂). ¹³C NMR (75 MHz, CDCl₃):168.17, 147.50, 145.10, 144.60, 136.91, 135.01, 133.07, 132.01, 131.49,130.25, 130.04, 128.73, 128.00, 126.83, 123.90, 123.35, 95.03, 59.23,58.51, 42.60. m/z (ES): 465 [M+H]⁺. HPLC: R_(t)=3.51 min. Anal.: calc.for C₂₅H₂₁ClN₂O₅: C, 64.59; H, 4.55; N, 6.02. Found: C, 64.23; H, 4.63;N, 5.73.

3-(4-Chlorophenyl)-3-(5-hydroxycyclooctyloxy)-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(NU8352)

The named compound was synthesised from3-(4-Chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(400 mg, 1.01 mmol, 1 equiv.) and cis-1,5-cyclooctanediol (728 mg, 5.05mmol, 5 equiv.) using General Procedure A and obtained as a yellow solid(342 mg, 65%). ¹H NMR (300 MHz, CDCl₃): 7.95-7.91 (m, 3H, O₂N—C—CH andC(O)═C═CH), 7.60-7.52 (m, 2H, Ar—H), 7.18-7.03 (m, 7H, Ar—H), 4.86 and4.20 (dd: AB, J=15.3 Hz, 2H, N—CH₂), 3.59-3.52 (m, 1H, HO—CH), 3.24-3.18(m, 1H, O—CH), 1.83-1.29 (m, 12H, CH₂). ¹³C NMR (75 MHz, CDCl₃): 168.29,147.21, 145.92, 144.85, 137.61, 134.77, 132.60, 131.79, 130.16, 129.68,128.34, 128.27, 124.02, 123.62, 123.08, 94.52, 73.62, 71.25, 42.75,36.51, 35.92, 34.32, 33.94, 20.30, 20.12 Anal.: calc. forC₃₉H₃₂ClN₃O₆+0.5 EtOAc: C, 66.34; H, 5.77; N, 5.16. Found: C, 66.25; H,5.91; N, 5.00. Mp: 69-72° C. (EtOAc).

3-(4-Chloro-phenyl)-3-(3-hydroxy-2,2-dimethyl-propoxy)-2-(4-nitro-benzyl)-2,3-dihydro-isoindol-1-one(NU8353)

The named compound was synthesised from3-(4-Chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(400 mg, 1.01 mmol, 1 equiv.) and neopentyl glycol (526 mg, 5.05 mmol, 5equiv.) using General Procedure A and obtained as an off-white solid(267 mg, 55%). ¹H NMR (300 MHz, CDCl₃): 8.02-7.98 (m, 2H, O₂N—C—CH),7.96-7.93 (m, 1H, C(O)═C═CH), 7.58-7.55 (m, 2H, Ar—H), 7.32-7.28 (m, 2H,Ar—H), 7.15-7.12 (m, 5H, Ar—H), 4.58 and 4.44 (dd: AB, J=15.3 Hz, 2H,N—CH₂), 3.39 (s, 2H, HO—CH₂), 2.78 and 2.63 (dd: AB, J=8.7 Hz, 2H,N—CH₂), 0.83 (d, J=3.9 Hz, CH₃). ¹³C NMR (75 MHz, CDCl₃): 168.30,147.33, 145.19, 144.60, 137.31, 134.90, 133.08, 131.64, 130.17, 129.85,128.61, 127.95, 123.81, 123.23, 123.16, 94.54, 69.57, 69.02, 42.42,36.41, 21.75. m/z (ES): 481 [M+H]⁺.

Anal.: calc. for C₂₆H₂₅ClN₂O₅+0.25H₂O: C, 64.32; H, 5.31; N, 5.77.Found: C, 64.32; H, 5.34; N, 5.57.

3-(4-Chlorophenyl)-3-(1-hydroxymethylcyclopropylmethoxy)-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(NU8354)

The named compound was synthesised from3-(4-Chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(400 mg, 1.01 mmol, 1 equiv.) and cyclopropane dimethanol (516 mg, 5.05mmol, 5 equiv.) using General Procedure A and obtained as an off-whitesolid (305 mg, 63%). ¹H NMR (300 MHz, CDCl₃): 8.01-7.98 (m, 2H,O₂N—C—CH), 7.92-7.89 (m, 1H, C(O)═C═CH), 7.55-7.52 (m, 2H, Ar—H),7.32-7.29 (m, 2H, Ar—H), 7.19-7.12 (m, 5H, Ar—H), 4.49 (s, 2H, N—CH₂),3.51-3.43 (m, 2H, HOCH₂), 2.81 (s, 2H, OCH₂), 0.43-0.40 (m, 2H, CH₂),0.22-0.12 (m, 2H, CH₂). ¹³C NMR (75 MHz, CDCl₃): 168.54, 145.52, 144.95,137.47, 135.29, 133.45, 131.87, 130.50, 130.21, 128.98, 128.30, 124.16,123.60, 123.48, 94.96, 67.84, 42.75, 22.68, 8.94, 8.90 Anal.: calc. forC₂₆H₂₃ClN₂O₅: C, 65.20; H, 4.84; N, 5.85, Found: C, 64.83; H, 4.92; N,5.63.

Racemic NU8354 was separated into its two enantiomers by chiral HPLC(Chiracel AD column; 1 cm×25 cm; 40% EtOH, pentane):NU8354A, RT=9.8 min; α=+22.66°, 0.406 g/100 ml; and NU8354B, RT=12.4min; α=−20.10°, 0.398 g/100 ml.

3-(4-Chlorophenyl)-3-(4-hydroxybut-2-ynyloxy)-2-(4-nitrobenzyl)-2,3dihydroisoindol-1-one (NU8357)

The named compound was synthesised from3-(4-Chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(400 mg, 1.01 mmol, 1 equiv.) and butynediol (435 mg, 5.05 mmol, 5equiv.) using General Procedure A and obtained as a yellow solid (271mg, 58%). ¹H NMR (300 MHz, CDCl₃): 8.05-7.93 (m, 3H, O₂N—C—CH andC(O)═C═CH), 7.61-7.54 (m, 2H, Ar—H), 7.36-7.33 (m, 2H, Ar—H), 7.24-7.16(m, 5H, Ar—H), 4.60 and 4.52 (d: AB, J=15.0 Hz, 2H, N—CH₂), 4.20-4.18(m, 2H, HO—CH₂), 3.83 and 3.52 (dt: AB, J=1.8, 15.3 Hz, 2H, O—CH₂), 2.42(t, J=6.0 Hz, 1H, OH). ¹³C NMR (75 MHz, CDCl₃): 169.24, 148.20, 145.18,137.24, 135.88, 133.78, 132.49, 131.29, 130.78, 129.43, 128.79, 124.81,124.43, 124.14, 95.88, 86.01, 81.28, 54.04, 52.97, 51.55, 43.60. m/z(ES): 463 [M+H]⁺. Anal.: calc. for C₂₅H₁₉ClN₂O₅+0.2H₂O: C, 64.36; H,4.20; N, 6.01. Found: C, 64.11; H, 3.72; N, 5.53.

3-(4-Chlorophenyl)-3-(4-hydroxymethylcyclohexylmethoxy)-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(NU8358)

The named compound was synthesised from3-(4-Chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(400 mg, 1.01 mmol, 1 equiv.) and trans-cyclohexane-1,4-dimethanol (728mg, 5.05 mmol, 5 equiv.) using General Procedure A and obtained as ayellow solid (374 mg, 71%). ¹H NMR (300 MHz, CDCl₃): 8.05-7.92 (m, 3H,O₂N—C—CH and C(O)═C═CH), 7.56-7.53 (m, 2H, Ar—H), 7.38-7.10 (m, 7H,Ar—H), 4.59 and 4.35 (d: AB, J=15.0 Hz, 2H, N—CH₂), 3.44 (d, J=6.3 Hz,2H, HO—CH₂), 2.65-2.53 (m, 2H, OCH₂), 1.81-1.74 (m, 3H, OH and CH),1.44-1.32 (m, 2H), 0.93-0.79 (m, 5H). ¹³C NMR (75 MHz, CDCl₃): 168.26,145.41, 144.82, 137.42, 134.84, 132.93, 130.01, 128.63, 128.01, 123.80,123.24, 123.18, 94.67, 68.41, 68.26, 42.41, 40.59, 38.04, 29.68, 29.31,28.90. m/z (ES): 521 [M+H]⁺. Anal.: calc. for C₂₉H₂₉ClN₂O₅+0.2H₂O: C,66.38; H, 5.66; N, 5.34. Found: C, 66.23; H, 5.79; N, 4.99.

3-(4-Chlorophenyl)-3-(2-hydroxymethylcyclohexylmethoxy)-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(NU8359)

The named compound was synthesised from3-(4-Chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(400 mg, 1.01 mmol, 1 equiv.) and cis-1,2-cyclohexane-dimethanol (728mg, 5.05 mmol, 5 equiv.) using General Procedure A and obtained as ayellow solid (337 mg, 64%). ¹H NMR (300 MHz, CDCl₃): mixture ofdiastereoisomers: 8.08-7.93 (m, 3H, O₂N—C—CH and C(O)═C═CH), 7.58-7.54(m, 2H, Ar—H), 7.43-7.36 (m, 2H, Ar—H), 7.23-7.11 (m, 5H, Ar—H), 4.61and 4.25 (d: AB, J=15.0 Hz, 2H, N—CH₂); and 4.59 and 4.36 (d: AB, J=15.0Hz, 2H, N—CH₂), 3.53-3.24 (m, 2H, HO—CH₂), 2.87-2.61 (m, 2H, OCH₂),1.87-1.09 (m, 10H, CH). ¹³C NMR (75 MHz, CDCl₃): mixture ofdiastereoisomers: 168.25, 147.42, 145.37, 145.20, 144.84, 144.76,137.28, 137.12, 134.97, 134.88, 133.03, 131.55, 131.41, 130.15, 130.10,129.98, 129.94, 128.81, 128.69, 127.98, 127.92, 123.91, 123.81, 123.33,123.30, 123.10, 123.06, 95.07, 94.97, 63.91, 63.57, 63.44, 63.31, 42.50,42.43, 40.77, 39.99, 37.18, 27.40, 27.09, 26.21, 26.16, 23.77, 23.44,23.29. Anal.: calc. for C₂₉H₂₉ClN₂O₅+0.3 CH₂Cl₂+0.1H₂O: C, 64.18; H,5.49; N, 5.11. Found: C, 64.34; H, 5.49; N, 4.95.

3-(4-Chlorophenyl)-3-(4-hydroxycyclohexyloxy)-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(NU8360)

The named compound was synthesised from3-(4-Chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(400 mg, 1.01 mmol, 1 equiv.) and cis/trans-1,4-cyclohexanediol (586 mg,5.05 mmol, 5 equiv.) using General Procedure A and obtained as a whitesolid (338 mg, 68%). ¹H NMR (300 MHz, CDCl₃): mixture ofdiastereoisomers: 7.93-7.90 (m, 3H, O₂N—C—CH and C(O)═C═CH), 7.56-7.51(m, 2H, Ar—H), 7.18-7.02 (m, 7H, Ar—H), 4.78 and 4.24 (d: AB, J=15.0 Hz,2H, N—CH₂); 3.70-3.62 (m, 1H, HOCH—), 3.26-3.09 (m, 1H, OCH), 1.86-1.26(m, 10H, CH₂). ¹³C NMR (75 MHz, CDCl₃): mixture of diastereoisomers:168.33, 147.16, 146.01, 144.80, 137.66, 134.74, 132.72, 131.58, 130.21,130.16, 129.68, 128.31, 128.28, 128.23, 128.19, 123.81, 123.09, 94.30,94.28, 71.41, 69.22, 68.61, 67.66, 42.65, 32.51, 32.32, 30.95, 30.65,30.33, 29.19, 29.03. m/z (ES): 493 [M+H]⁺. HPLC: R_(t)=3.43 min Anal.:calc. for C₂₇H₂₅ClN₂O₅+0.1H₂O: C, 65.54; H, 5.14; N, 5.66. Found: C,65.09; H, 5.20; N, 5.24.

3-(4-Chlorophenyl)-3-(4-hydroxycyclohex-2-enyloxy)-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(NU8361)

The named compound was synthesised from3-(4-chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(400 mg, 1.01 mmol, 1 equiv.) and trans-1,4-cyclohex-2-enediol (576 mg,5.05 mmol, 5 equiv.) using General Procedure A and obtained as a whitesolid (263 mg, 53%). ¹H NMR (300 MHz, CDCl₃): mixture ofdiastereoisomers: 7.96-7.92 (m, 3H, O₂N—C—CH and C(O)═C═CH), 7.61-7.55(m, 2H, Ar—H), 7.22-7.03 (m, 7H, Ar—H), 5.75 and 5.34 (m, 2H, CH═CH),4.83 and 4.28 (m, 2H, N—CH₂), 4.27 (m, 1H, HO—CH), 3.74-3.71 (m, 1H,OCH), 3.70-3.62 (m, 1H, HOCH—), 3.26-3.09 (m, 1H, OCH), 2.12-1.14 (m,4H, CH₂). ¹³C NMR (75 MHz, CDCl₃): mixture of diastereoisomers: 168.57,147.55, 146.08, 145.06, 144.99, 137.66, 137.57, 135.16, 134.64, 134.10,133.20, 131.90, 130.68, 130.61, 130.09, 130.04, 129.96, 128.67, 128.65,128.59, 128.55, 124.28, 124.15, 123.48, 123.46, 94.90, 94.82, 68.43,68.30, 65.91, 65.76, 43.06, 30.85, 30.52, 28.58. m/z (ES): 491 [M+H]⁺.HPLC: R_(t)=3.39 min Anal.: calc. for C₂₇H₂₃ClN₂O₅+0.2H₂O: C, 65.57; H,4.78; N, 5.67. Found: C, 65.32; H, 5.00; N, 5.18.

3-(4-Chlorophenyl)-3-hydroxy-2-[2-(4-nitrophenyl)ethyl]-2,3-dihydroisoindol-1-one

To a solution of 2-(4-chlorobenzoyl)benzoic acid (5 g, 19.2 mmol, 1equiv.) in dry THF (20 mL) was added under nitrogen atmospherethionylchloride (3.0 mL, 38.3 mmol, 2 equiv.) and 3 drops of anhydrousDMF. The reaction mixture was stirred for 4 h at room temperature andconcentrated in vacuo. The resulting pale yellow oil was taken up in dryTHF (20 mL), and the amine 2-(4-nitrophenyl)ethylamine hydrochloride(4.30 g, 21.1 mmol, 1.1 equiv.) and DIPEA (3.49 mL, 21.1 mmol, 1.1equiv.) were added under nitrogen atmosphere. The reaction mixture wasstirred overnight at room temperature and the solvents were removed invacuo. The residue was taken up in EtOAc (100 mL), filtered, and thefiltrate washed with water (3×50 mL) and brine (1×50 mL). The organiclayer was dried over MgSO₄, filtered and concentrated in vacuo to afforda buff-coloured solid, which was recrystallised from EtOAc/Petrol ether(4.47 g, 57%). ¹H NMR (300 MHz, d⁶-DMSO): 8.09 (d, J=8.4 Hz, 2H,O₂N—C—CH), 7.74 (d, J=6.6 Hz, 1H, C(O)═C═CH), 7.60-7.51 (m, 2H, Ar—H),7.40-7.26 (m, 8H, Ar—H and OH), 3.69-3.57 (m, 1H, N—CH), 3.28-3.19 (m,1H, N—CH), 2.96-2.89 (m, 2H, N—CH₂—CH₂). ¹³C NMR (75 MHz, d⁶-DMSO):166.38, 148.77, 147.11, 145.85, 138.81, 132.57, 132.21, 130.08, 129.34,128.97, 128.05, 127.51, 122.95, 122.36, 122.13. FTIR: 3240, 1166, 1520,1338 cm⁻¹. m/z (ES): 409 [M+H]⁺. HPLC: R_(t)=3.36 min. Anal.: calc. forC₂₂H₁₇ClN₂O₄: C, 64.63, H, 4.19, N, 6.85. Found: C, 65.02; H, 4.27; N,6.74. Mp: 206-208° C. (EtOAc). UV: λ_(max)=268 nm (EtOH).

3-(4-Chlorophenyl)-3-(4-hydroxybutoxy)-2-[2-(4-nitrophenyl)ethyl]-2,3-dihydroisoindol-1-one(NU8362)

The named compound was synthesised from3-(4-chlorophenyl)-3-hydroxy-2-[2-(4-nitrophenyl)ethyl]-2,3-dihydroisoindol-1-one(562 mg, 1.37 mmol, 1 equiv.) and 1,4-butanediol (616 mg, 6.85 mmol, 5equiv.) using General Procedure A and obtained as a white solid (306 mg,63%). ¹H NMR (300 MHz, CDCl₃): 8.11-8.08 (m, 2H, O₂N—C—CH), 7.91-7.88(m, 1H, C(O)═C═CH), 7.55-7.53 (m, 2H, Ar—H), 7.30-7.14 (m, 7H, Ar—H),3.66 (m, 2H, N—CH₂), 3.45 (t, J=8.1 Hz, 2H, HO—CH₂), 3.16-3.11 and3.03-2.99 (m, 2H, OCH₂), 3.00-2.89 and 2.73-2.66 (m, 2H, NCH₂CH₂),1.75-1.63 (m, 5H, CH₂ and OH). ¹³C NMR (75 MHz, CDCl₃): 168.23, 147.02,146.62, 145.32, 137.69, 134.84, 132.74, 131.98, 129.99, 129.48, 128.74,127.87, 123.65, 123.51, 123.10, 94.54, 62.79, 62.43, 40.51, 34.21,29.60, 26.07. HPLC: R_(t)=3.45 min. Anal.: calc. for C₂₆H₂₅ClN₂O₅: C,64.93; H, 5.24; N, 5.82. Found: C, 64.82; H, 5.18; N, 5.68.

3-(4-Chlorophenyl)-2-[1-(4-chlorophenyl)-ethyl]-3-(4-hydroxybutoxy)-2,3-dihydroisoindol-1-one(NU8365)

The named compound was synthesised from3-(4-chlorophenyl)-2-[1-(4-chlorophenyl)-ethyl]-3-hydroxy-2,3-dihydroisoindol-1-one(498 mg, 1.37 mmol, 1 equiv.) and 1,4-butanediol (616 mg, 6.85 mmol, 5equiv.) using General Procedure A and obtained as a pale yellow oil (242mg, 51%). ¹H NMR (300 MHz, CDCl₃): 7.87-7.83 (m, 1H, C(O)═C═CH),7.53-7.46 (m, 2H, Ar—H), 7.07-7.03 (m, 9H, Ar—H), 4.41 (q, J=7.2 Hz, 1H,N—CH), 3.70 (m, 2H, HO—CH₂), 3.29-3.25 and 3.01-2.95 (m, 2H, OCH₂), 1.86(d, J=7.2 Hz, 3H, CH₃), 1.77-1.68 (m, 4H, CH₂). ¹³C NMR (75 MHz, CDCl₃):168.31, 145.26, 141.47, 137.76, 134.79, 133.15, 133.06, 132.84, 130.20,129.67, 128.75, 128.41, 128.27, 123.71, 123.27, 95.44, 63.35, 62.89,52.49, 29.96, 26.48, 20.08. m/z (ES): 470 [M+H]⁺. HPLC: R_(t)=3.76 min.Anal.: calc. for C₂₆H₂₅Cl₂NO₃: C, 66.39; H, 5.36; N, 2.98. Found: C,66.33; H, 5.25; N, 2.46.

3-(4-Chlorophenyl)-3-(4-hydroxymethylbenzyloxy)-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(NU8366)

The named compound was synthesised from3-(4-chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(541 mg, 1.37 mmol, 1 equiv.) and 1,4-benzenedimethanol (946 mg, 6.85mmol, 5 equiv.) using General Procedure A and obtained as a white solid(405 mg, 78%). ¹H NMR (300 MHz, CDCl₃): 8.01-7.85 (m, 3H, O₂N—C—CH andC(O)═C═CH), 7.61-7.53 (m, 2H, Ar—H), 7.37-7.18 (m, 9H, Ar—H), 6.93 (d,J=7.8 Hz, 2H, Ar—H), 4.85 and 4.67 (d: AB, J=15.0 Hz, 2H, N—CH₂), 4.66(d, J=5.7 Hz, CH₂—OH), 3.90 and 3.68 (d: AB, J=11.7 Hz, 2H, OCH₂), 2.12(t, J=6.0 Hz, 1H, OH). ¹³C NMR (75 MHz, CDCl₃): 168.52, 147.63, 145.39,144.90, 141.16, 137.34, 136.39, 135.37, 133.42, 131.73, 130.57, 130.32,129.19, 128.38, 127.30, 127.11, 124.34, 123.69, 95.52, 65.26, 65.12,42.89. m/z (ES): 515 [M+H]⁺. HPLC: R_(t)=3.56 min. Anal.: calc. forC₂₉H₂₃ClN₂O₅: C, 67.64; H, 4.50; N, 5.44. Found: C, 67.38; H, 4.43; N,5.26.

3-(4-Chlorophenyl)-3-(3-hydroxymethylbenzyloxy)-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(NU8367)

The named compound was synthesised from3-(4-chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(541 mg, 1.37 mmol, 1 equiv.) and 1,3-benzenedimethanol (946 mg, 6.85mmol, 5 equiv.) using General Procedure A and obtained as a white solid(390 mg, 75%). ¹H NMR (300 MHz, CDCl₃): 8.01-7.90 (m, 3H, O₂N—C—CH andC(O)═C═CH), 7.62-7.53 (m, 2H, Ar—H), 7.39-7.17 (m, 9H, Ar—H), 6.95-6.91(m, 2H, Ar—H), 4.80 and 4.13 (d: AB, J=15.0 Hz, 2H, N—CH₂), 4.64 (d,J=5.1 Hz, CH₂—OH), 3.93 and 3.73 (d: AB, J=11.4 Hz, 2H, OCH₂), 1.95 (t,J=5.4 Hz, 1H, OH). ¹³C NMR (75 MHz, CDCl₃): 168.51, 145.42, 144.88,141.59, 137.45, 135.37, 133.39, 131.79, 130.56, 130.34, 129.17, 128.81,128.38, 126.61, 126.51, 125.82, 124.32, 123.69, 95.55, 65.42, 65.35,42.91. m/z (ES): 515 [M+H]⁺. HPLC: R_(t)=3.58 min. Anal.: calc. forC₂₉H₂₃ClN₂O₅+0.15 CH₂Cl₂: C, 66.34; H, 4.46; N, 5.31. Found: C, 66.12;H, 4.44; N, 5.03.

3-(4-Chloro-phenyl)-2-[1-(4-chloro-phenyl)-ethyl]-3-hydroxy-2,3-dihydro-isoindol-1-one

To a solution of (R)-4-chloro-α-methylbenzylamine hydrochloride (2.95mL, 21.1 mmol, 1.1 equiv.) in dry THF (20 mL) was added under nitrogenatmosphere thionylchloride (3.0 mL, 38.3 mmol, 2 equiv.) and 3 drops ofanhydrous DMF. The reaction mixture was stirred for 4 h at roomtemperature and concentrated in vacuo. The resulting pale yellow oil wastaken up in dry THF (20 mL), and the amine 2-(4-nitrophenyl)ethylaminehydrochloride (4.30 g, 21.1 mmol, 1.1 equiv.) and DIPEA (3.49 mL, 21.1mmol, 1.1 equiv.) were added under nitrogen atmosphere. The reactionmixture was stirred overnight at room temperature and the solvents wereremoved in vacuo. The residue was taken up in EtOAc (100 mL), filtered,and the filtrate washed with water (3×50 mL) and brine (1×50 mL). Theorganic layer was dried over MgSO₄, filtered and concentrated in vacuoto afford a white crystalline powder, which was recrystallised fromEtOAc/Petrol ether (4.98 g, 65%). ¹H NMR (300 MHz, d⁶-DMSO): Mixture oftwo diastereoisomers: 7.64-7.61 (m, 1H and 1H, Ar—H), 7.46-7.30 (m, 5Hand 5H, Ar—H), 7.26-7.02 (m, 5H and 5H, Ar—H), 6.90 (d, J=8.7 Hz, 1H and1H, Ar—H), 4.58 and 4.43 (q, J=6.9 Hz, 1H, CH*), 4.18 and 4.11 (br s,1H, OH), 1.69 and 1.56 (d, J=7.2 Hz, 3H, CH₃). ¹³C NMR (75 MHz,d⁶-DMSO): 167.33, 167.26, 148.29, 148.25, 141.08, 140.53, 137.44,137.32, 134.74, 134.67, 132.96, 132.91, 132.75, 132.72, 131.30, 131.21,129.82, 129.45, 129.40, 128.81, 128.70, 128.43, 128.36, 128.25, 128.00,123.44, 123.41, 122.47, 91.93, 91.50, 52.07, 52.01, 19.84, 18.43. FTIR:3119, 1667 cm⁻¹. m/z (ES): 398 [M+H]⁺. HPLC: R_(t)=3.57 min. Anal.:calc. for C₂₂H₁₇Cl₂NO₂: C, 66.34; H, 4.30; N, 3.52. Found: C, 66.24; H,4.28; N, 3.50.

3-(4-Chloro-phenyl)-2-[1-(4-chloro-phenyl)-ethyl]-3-(4-hydroxy-butoxy)-2,3-dihydro-isoindol-1-one(NU8368)

The named compound was synthesised from3-(4-chloro-phenyl)-2-[1-(4-chloro-phenyl)-ethyl]-3-hydroxy-2,3-dihydro-isoindol-1-one(498 mg, 1.37 mmol, 1 equiv.) and 1,4-butanediol (616 mg, 6.85 mmol, 5equiv.) using General Procedure B and obtained as a clear oil (304 mg,64%). ¹H NMR (300 MHz, CDCl₃): 7.87-7.84 (m, 1H, C(O)═C═CH), 7.53-7.46(m, 2H, Ar—H), 7.07-7.00 (m, 9H, Ar—H), 4.41 (q, J=7.2 Hz, 1H, N—CH),3.70 (m, 2H, HO—CH₂), 3.31-3.24 and 3.02-2.95 (m, 2H, OCH₂), 1.86 (d,J=7.2 Hz, 3H, CH₃), 1.76-1.66 (m, 4H, CH₂). ¹³C NMR (75 MHz, CDCl₃):168.32, 145.27, 141.48, 137.77, 134.80, 133.15, 133.06, 132.84, 130.20,129.67, 128.74, 128.41, 123.72, 123.27, 95.45, 63.36, 62.88, 52.49,29.96, 26.48, 20.08. m/z (ES): 470 [M+H]⁺. Anal.: calc. forC₂₆H₂₅Cl₂NO₃+0.3H₂O: C, 65.62; H, 5.43; N, 2.94. Found: C, 65.58; H,5.77; N, 2.45.

4-[1-(4-Chloro-phenyl)-1-(4-hydroxy-butoxy)-3-oxo-1,3-dihydro-isoindol-2-ylmethyl]-benzonitrile(NU8370)

The named compound was synthesised from2-(4-(aminomethyl)benzonitrile)-3-(4-chlorophenyl)-3-hydroxyisoindolin-1-one(513 mg, 1.37 mmol, 1 equiv.) and 1,4-butanediol (616 mg, 6.85 mmol, 5equiv.) using General Procedure B and obtained as a white solid (302 mg,67%). ¹H NMR (300 MHz, CDCl₃): 7.94-7.91 (m, 1H, C(O)═C═CH), 7.55-7.52(m, 2H, Ar—H), 7.47 and 7.33 (d: A₂B₂, J=8.4 Hz, 4H, Ar—H), 7.20 (m, 4H,Ar—H), 7.13-7.10 (m, 1H, Ar—H), 4.60 and 4.24 (d: AB, J=15.0 Hz, 2H,NCH₂), 3.56 (t, J=5.4 Hz, 2H, HOCH₂), 2.80-2.74 (m, 2H, OCH₂), 1.53-1.16(m, 4H, CH₂). ¹³C NMR (75 MHz, CDCl₃): 168.18, 145.32, 142.86, 137.20,134.77, 132.90, 131.86, 131.46, 129.99, 129.85, 128.61, 127.95, 123.77,123.08, 118.41, 111.33, 94.78, 62.82, 62.34, 42.68, 29.41, 25.67. Anal.:calc. for C₂₉H₂₃ClN₂O₃+0.2H₂O: C, 69.37; H, 5.16; N, 6.22. Found: C,69.29; H, 5.20, N: 6.04.

All references to General Procedures A-F made hereinbelow are referencesto the General Procedures A-F outlined immediately below and do notrefer to General Procedures A-C hereinabove.

General Procedure A

To a suspension of the corresponding phthalic anhydride (1 equiv.) inchlorobenzene (8 equiv.) was added aluminium chloride (2.4 equiv.) Themixture was heated to 90° C. for 2 h and then cooled to roomtemperature. Ice was added followed by conc. HCl (5 mL) and the mixturewas extracted into dichloromethane (DCM) (3×50 mL) and then washed with10% Na₂CO₃ solution (2×50 mL). The Na₂CO₃ washings were combined andacidified to pH3 with conc. HCl. The resulting precipitate was collectedby filtration and dried in a vacuum oven.

General Procedure B

To a solution of the corresponding benzoic acid (1 equiv.) in dry THF(10 mL) was added under a nitrogen atmosphere thionyl chloride (2equiv.) and 3 drops of anhydrous DMF. The reaction mixture was stirredfor 4 h at room temperature and concentrated in vacuo. The resultingpale yellow oil was taken up in dry THF (10 mL) and the amine (1.1equiv.) and DIPEA (1.1 equiv.) were added under nitrogen atmosphere. Thereaction mixture was stirred overnight at room temperature and thesolvents were removed in vacuo. The residue was taken up in EtOAc (50mL), filtered and the filtrate washed with water (3×25 mL) and brine(1×25 mL). The organic layer was dried over MgSO₄, filtered andconcentrated in vacuo to afford a solid which was recrystallised fromEtOAc/petrol ether or purified by flash chromatography (Biotage SP4).

General Procedure C

To solution of the corresponding isoindolinone (1 equiv.) in dry THF (10mL) was added under a nitrogen atmosphere thionyl chloride (2 equiv.)and 3 drops of anhydrous DMF. The reaction mixture was stirred for 4 hat room temperature and concentrated in vacuo. The resulting pale yellowoil was taken up in dry THF (10 mL) and the alcohol (2 equiv.) andpotassium carbonate (2 equiv.) were added. The reaction mixture wasstirred overnight at room temperature and the solvents were removed invacuo. The residue was taken up in EtOAc (50 mL) and washed with water(3×25 mL) and brine (1×25 mL). The organic layer was dried over MgSO₄,filtered and concentrated in vacuo to afford an oil which was purifiedby flash chromatography (Biotage SP4).

General Procedure D

To a solution of the corresponding isoindolinone (1 equiv.) in anhydrousDCM (5 mL) was added mCPBA (1.1 equiv.). The reaction mixture wasstirred at 30° C. for 4 h and then diluted with DCM (30 mL), washed withsaturated NaHCO₃ solution (30 mL), water (30 mL) and brine (30 mL). Theorganic layer was dried over MgSO₄, filtered and concentrated in vacuoto afford an oil which was purified by flash chromatography (BiotageSP4).

General Procedure E

To a solution of the corresponding ester (1 equiv.) in dry THF was addedunder a nitrogen atmosphere potassium trimethylsilanolate (1.1 equiv.).The reaction mixture was stirred at room temperature overnight. Furtherpotassium trimethylsilanolate (1.1 equiv.) was added and the mixture wasagain stirred at room temperature overnight. The solvent wasconcentrated in vacuo to afford a solid which was purified by flashchromatography (Biotage SP4).

General Procedure F

To a solution of the corresponding isoindolinone (1 equiv.) in THF (10mL) was added pyridine (2 equiv.), 4-dimethylamino pyridine (catalytic)and succinic anhydride (2 equiv.). The reaction mixture was heated underreflux for 48 h, cooled to RT and the solvent concentrated in vacuo. Theresidue was dissolved in EtOAc (50 mL), washed with water (2×20 mL),brine (20 mL), dried over Na₂SO₄ and concentrated in vacuo. The productwas purified by flash chromatography.

INTERMEDIATES Synthesis of 2-(4-bromobenzoyl)benzoic acid

To a suspension of the phthalic anhydride (2 g, 13.50 mmol) inbromobenzene (11.38 mL, 108 mmol) was added aluminium chloride (3.60 g,27.00 mmol). The mixture was heated to 90° C. for 2 h and then cooled toroom temperature. Ice was added followed by conc. HCl (5 mL) and themixture was extracted into DCM (3×50 mL) and then washed with 10% Na₂CO₃solution (2×50 mL). The Na₂CO₃ washings were combined and acidified topH3 with conc. HCl. The resulting precipitate was collected byfiltration and dried in a vacuum oven. The named compound was obtainedas a white solid (3.41 g, 83%).

¹H NMR (300 MHz, DMSO) δ 7.39-7.42 (m, 1H, —CH═CH—C(CO₂H)), 7.51-7.53 (dAB, J=7.7 Hz, 2H, —CH—C(Br)), 7.73-7.66 (m, 4H, ArH), 7.97-8.00 (m, 1H,—CH═CH—CH—C(CO₂H))

¹³C NMR (DMSO, 75 MHz), δ 127.61, 128.89, 129.97, 130.09, 130.38,130.71, 130.92, 132.07, 132.96, 136.46, 167.09, 194.21

IR: 665, 702, 736, 770, 812, 839, 924, 1009, 1065, 1148, 1252, 1279,1422, 1485, 1570, 1670, 2546, 2657, 2832, 2988 cm⁻¹

LCMS (DMSO): Rt=3.01 min (on 5 min column)

UV (in EtOH): λ max=259 nm

Rf: 0.29 (50% EtOAc/petrol)

MP: 170-172° C.

Synthesis of a mixture of 2-(4-chlorobenzoyl)-3-methylbenzoic acid and2-(4-chlorobenzoyl)-6-methylbenzoic acid

The named compounds were synthesised from 3-methylphthalic anhydride (3g, 18.50 mmol) using General Procedure A and obtained as a white solid(2.98 g, 59%, ratio of 3- and 6-isomers is 20:1) which was used withoutfurther purification.

¹H NMR (300 MHz, DMSO)

Major isomer: δ 2.06 (s, 3H, CH ₃), 7.45-7.61 (m, 6H, ArH), 7.86-7.88(d, J=6.6 Hz, 1H, CH—C(CO₂H)), 10.69 (br s, 1H, CO₂ H)

¹³C NMR (DMSO, 75 MHz), δ 18.63, 127.36, 128.94, 129.22, 129.53, 129.65,130.06, 132.60, 134.94, 135.23, 137.74, 167.31, 193.10

IR: 675, 738, 754, 831, 919, 1009, 1088, 1144, 1264, 1288, 1400, 1580,1678, 1749, 2556, 2643, 2817, 2961, 3406 cm⁻¹

LCMS (DMSO): Rt=3.49 min (on 5 min column)

UV (in EtOH): λ max=255 nm

Rf: 0.37 (50% EtOAc/petrol)

MP: 179-180° C.

Synthesis of a mixture of 2-(4-chlorobenzoyl)-4-methylbenzoic acid and2-(4-chlorobenzoyl)-5-methylbenzoic acid

The named compounds were synthesised from 4-methylphthalic anhydride (3g, 18.50 mmol) using General Procedure A and obtained as a white solid(4.60 g, 90%, ratio of 4- and 5-isomers is 2:1) which was used withoutfurther purification.

¹H NMR (300 MHz, DMSO) δ

Major isomer: 2.41 (s, 3H, CH ₃), 7.23-7.25 (d, J=1.4 Hz, 1H,C(Me)—CH—C(COAr)), 7.43-7.48 (m, 1H, C(Me)-CH—CH—C(CO₂H)), 7.53-7.63 (m,4H, ArH), 7.89-7.91 (d, J=8.0 Hz, 1H, —CH—CH—C(CO₂H)), 13.10 (br s, 1H,CO₂ H)

Minor isomer: 2.44 (s, 3H, CH ₃), 7.32-7.34 (d, J=7.7 Hz, 1H,C(Me)-CH—CH—C(COAr), 7.36-7.40 (m, 1H, C(Me)-CH—CH—C(COAr), 7.53-7.63(m, 4H, ArH), 7.80-7.82 (d, J=1.1 Hz, 1H, C(Me)-CH—C(CO₂H)), 13.10 (brs, 1H, CO₂ H)

¹³C NMR (DMSO, 75 MHz), δ 21.28, 127.79, 127.95, 129.07, 130.12, 130.78,133.17, 136.45, 137.22, 138.11, 140.24, 167.00, 190.90

IR: 683, 751, 780, 838, 934, 962, 1009, 1088, 1153, 1211, 1288, 1398,1422, 1486, 1570, 1677, 2164, 2828, 3062 cm⁻¹

LCMS (DMSO): Rt=3.24 min (on 5 min column)

UV (in EtOH): λ max=253 nm

Rf: 0.42 (50% EtOAc/petrol)

MP: 163-164° C.

Synthesis of a mixture of 4-tert-butyl-2-(4-chlorobenzoyl)benzoic acidand 5-tert-butyl-2-(4-chlorobenzoyl)benzoic acid

The named compounds were synthesised from 4-tertbutylphthalic anhydride(2.5 g, 12.24 mmol) using General Procedure A and obtained as a creamsolid (3.56 g, 92%, ratio of 4- and 5-isomers is 1.3:1) which was usedwithout further purification.

¹H NMR (300 MHz, DMSO) δ

Major isomer: 1.29 (s, 9H, (CH ₃)₃), 7.35-7.38 (d, J=1.8 Hz, 1H,C(^(t)Bu)-CH—CH—C(CO₂H)), 7.54-7.69 (m, 4H, ArH), 7.69-7.75 (m, 1H,C(^(t)Bu)-CH—C(COAr)), 7.94-7.97 (d, J=8.3 Hz, 1H, —CH—CH—C(CO₂H))

Minor isomer: 1.34 (s, 9H, (CH ₃)₃), 7.40-7.41 (d, J=8.0 Hz, 1H,C(^(t)Bu)-CH—CH—C(COAr), 7.54-7.69 (m, 4H, ArH), 7.74-7.77 (m, 1H,C(^(t)Bu)-CH—CH—C(COAr), 7.99-8.00 (d, J=1.9 Hz, 1H,C(^(t)Bu)-CH—C(CO₂H))

Both isomers:

¹³C NMR (DMSO, 75 MHz), δ 31.07, 31.18, 34.96, 35.24, 124.45, 126.68,126.99, 127.59, 127.85, 128.69, 129.06, 129.56, 130.22, 130.52, 130.87,130.92, 136.41, 138.27, 138.61, 141.35, 153.12, 156.11, 166.93, 167.41,195.70, 195.88

IR: 683, 712, 762, 808, 843, 907, 932, 1007, 1083, 1119, 1159, 1252,1280, 1364, 1398, 1417, 1476, 1585, 2869, 2964 cm⁻¹

LCMS (DMSO): Rt=3.56 min (on 5 min column)

UV (in EtOH): λ max=255 nm

Rf=0.52 (50% EtOAc/petrol).

MP: 173-174° C.

Synthesis of 3-chloro-2-(4-chlorobenzoyl)benzoic acid and6-chloro-2-(4-chlorobenzoyl)benzoic acid

The named compounds were synthesised from 3-chlorophthalic anhydride (5g, 27.39 mmol) using General Procedure A and obtained as a yellow solid(6.71 g, 83%, ratio of 3- and 6-isomers is 99:1) which was used withoutfurther purification.

¹H NMR (300 MHz, DMSO) δ

Major isomer (20): 7.56-7.59 (d AB, J=8.6 Hz, CH—C(Cl)—CH), 7.65-7.71(m, 3H, ArH), 7.84-7.87 (dd, J=0.9, 8.0 Hz, 1H, CH—C(Cl)—C(COAr)),8.03-8.06 (dd, J=0.9, 8.0 Hz, 1H, CH—C(CO₂H)), 13.65 (br s, 1H, CO₂H)

Minor isomer (21): too weak to analyse

¹³C NMR (DMSO, 75 MHz), δ 129.38, 130.51, 130.59, 130.64, 131.36,131.82, 134.20, 135.73, 138.55, 139.88, 165.93, 195.17

IR: 672, 714, 743, 756, 824, 864, 920, 1009, 1090, 1154, 1206, 1255,1298, 1400, 1461, 1582, 1676, 2656, 2825, 3067 cm⁻¹

LCMS (DMSO): Rt=3.34 min (on 5 min column)

UV (in EtOH): λ max=256 nm

Rf=0.29 (50% EtOAc/petrol)

MP: 184-185° C.

Synthesis of 4-bromo-2-(4-chlorobenzoyl)benzoic acid and5-bromo-2-(4-chlorobenzoyl)benzoic acid

The named compounds were synthesised from 4-tertbutylphthalic anhydride(2.5 g, 12.24 mmol) using General Procedure A and obtained as a creamsolid (3.47 g, 66%, ratio of 4- and 5-isomers is ˜1:1) which was usedwithout further purification.

¹H NMR (300 MHz, DMSO) 6

Compound 36: 7.54-7.66 (m, 4H, —C₆ H ₄Cl), 7.84-7.93 (m, 2H, ArH),8.08-8.09 (d. J=1.7 Hz, 1H, C(Br)CHC(CO₂H)), 13.60 (br s, 1H, CO₂ H)

Compound 35: 7.54-7.66 (m, 4H, —C₆ H ₄Cl), 7.70-7.72 (d, J=1.5 Hz, 1H,C(Br)CHC(COAr)), 7.84-7.93 (m, 2H, ArH), 13.60 (br s, 1H, CO₂ H)

¹³C NMR (DMSO, 75 MHz), δ 128.16, 129.15, 130.21, 130.90, 131.37,132.12, 133.18, 135.21, 135.52, 138. 35, 170.54, 193.35

IR: 756, 841, 883, 927, 1013, 1092, 1152, 1177, 1285, 1400, 1423, 1483,1580, 1672, 2546 cm⁻¹

LCMS (DMSO): Rt=3.43 min (on 5 min column)

UV (in EtOH): λ max=255 nm

Rf=0.24 (50% EtOAc/petrol)

MP: 210-211° C.

Synthesis of 4,5-dichloro-2-(4-chlorobenzoyl)benzoic acid

The named compound was synthesised from 4,5-dichlorophthalic anhydride(2.5 g, 11.52 mmol) using General Procedure A and obtained as a yellowsolid (1.98 g, 52%).

¹H NMR (300 MHz, DMSO) δ 7.56-7.59 (d AB, J=8.3 Hz, —CC₂ H ₂C₂H₂Cl),7.67-7.70 (d AB, J=8.3 Hz, —CC₂ H ₂C₂H₂Cl), 7.87 (s, 1H, CH—C(COAr)),8.14 (s, 1H, CH—C(CO₂ H)), 13.85 (br s, 1H, CO₂ H)

¹³C NMR (DMSO, 75 MHz), δ 129.21, 130.76, 131.07, 131.11, 131.97,134.15, 137.45, 138.23, 138.70, 139.76, 170.36, 198.58

IR: 768, 843, 868, 893, 964, 1009, 1090, 1126, 1167, 1257, 1343, 1422,1478, 1545, 1582, 1678, 2228, 2563, 2834 cm⁻¹

UV (in EtOH): λ max=254 nm

Rf=0.18 (50% EtOAc/petrol)

MP: 195-197° C.

Synthesis of a mixture of 2-(4-chlorobenzoyl)-4-fluorobenzoic acid and2-(4-chlorobenzoyl)-5-fluorobenzoic acid

The named compounds were synthesised from 4-fluorophthalic anhydride(2.5 g, 15.05 mmol) using General Procedure A and obtained as a creamsolid (3.51 g, 84%, ratio of 4- and 5-isomers is 3:2). The mixture wasused without further purification.

¹H NMR (300 MHz, DMSO) 6

Major isomer: 7.24-7.27 (dd, J=2.0, 8.5 Hz, 1H, C(F)—CH—C(COAr)),7.40-7.46 (m, 1H, C(F)—CH—CH—C(CO₂H)), 7.49-7.52 (m, 2H, ArH), 7.57-7.61(m, 2H, ArH), 8.02-8.07 (dd, J=5.4, 8.5 Hz, 1H, CH—CH—(CO₂H))

Minor isomer: 7.40-7.46 (m, 1H, C(F)—CH—CH—C(COAr)), 7.49-7.52 (m, 3H,ArH), 7.57-7.61 (m, 2H, ArH), 7.66-7.70 (dd, J=1.9, 9.2 Hz, 1H,C(F)—CH—(CO₂H))

¹³C NMR (DMSO, 75 MHz), δ 116.98, 119.25, 128.62, 129.15, 130.85,130.92, 136.01, 138.40, 139.09, 166.10, 166.26, 196.59

IR: 681, 753, 784, 841, 860, 945, 974, 1011, 1086, 1140, 1174, 1211,1278, 1401, 1427, 1486, 1580, 1672, 2822, 3070 cm⁻¹

LCMS (DMSO): Rt=3.76 min (on 5 min column)

UV (in EtOH): λ max=256 nm

Rf=0.33 (50% EtOAc/petrol)

MP: 153-155° C.

Synthesis of 2-(4-chlorobenzoyl)cyclohex-1-enecarboxylic acid

The named compound was synthesised from 3,4,5,6-tetrahydrophthalicanhydride (2.08 g, 13.69 mmol) using General Procedure A, purified bychromatography (Biotage SP4; 20%-40% EtOAc/petrol) and obtained as ayellow oil (2.69 g, 74%).

¹H NMR (300 MHz, DMSO) δ 1.50-1.70 (m, 4H, CH ₂—C(COAr)—C(CO₂H)—CH ₂),2.15-2.17 (m, 4H, CH ₂—CH ₂—CH₂—C(CO₂H)), 7.93-7.48 (m, 4H, ArH), 8.16(br s, 1H, CO₂H)

¹³C NMR (DMSO, 75 MHz), δ 19.79, 21.01, 21.37, 21.45, 127.79, 128.84,133.95, 137.48, 146.33, 149.31, 171.24, 192.87

IR: 687, 729, 757, 819, 907, 938, 966, 1038, 1072, 1169, 1206, 1246,1375, 1422, 1489, 1597, 1736, 2865, 2937, 3318 cm⁻¹

LCMS (DMSO): Rt=3.72 min (on 5 min column)

UV (in EtOH): λ max=254 nm

Rf=0.48 (50% EtOAc/petrol)

Final Compounds Synthesis of3-(4-bromophenyl)-3-(4-hydroxybutoxy)-2-(4-nitrobenzyl)isoindolin-1-one(NU8390)

The named compound was synthesised from NU8389 (0.50 g, 1.14 mmol) and1,4-butanediol (0.20 mL, 2.28 mmol) using General Procedure C,purification by chromatography (Biotage SP4; 10%-50% EtOAc/petrol) andobtained as a pale yellow oil (0.50 g, 85%).

¹H NMR (300 MHz, CDCl₃) δ 1.36-1.45 (m, 4H, OCH₂—CH₂—CH₂), 2.76 (t,J=6.1 Hz, O—CH₂), 3.49 (t, J=6.2 Hz, HO—CH₂), 4.25 and 4.60 (dd: AB,J=15.0 Hz, 2H, N—CH₂), 7.08-7.13 (m, 3H, Ar—H), 7.29-7.36 (m, 4H, Ar—H),7.48-7.51 (m, 2H, Ar—H), 7.87-7.89 (m, 1H, C(O)═C═CH), 7.97-8.00 (m, 2H,O₂N—C—CH)

¹³C NMR (CDCl₃, 75 MHz), δ 26.01, 29.69, 42.69, 62.51, 63.19, 95.14,123.21, 123.45, 123.55, 124.09, 128.61, 130.25, 130.34, 131.76, 131.92,133.27, 138.13, 145.10, 145.58, 147.72, 168.49

IR: 696, 760, 804, 853, 928, 1005, 1063, 1099, 1177, 1276, 1341, 1381,1425, 1467, 1519, 1605, 1687, 2872, 2925, 3397 cm⁻¹

LCMS (DMSO): Rt=3.72 min (on 5 min column)

UV (in EtOH): λ max=267 nm

Rf: 0.26 (50% EtOAc/petrol)

CHN: C₂₅H₂₃BrN₂O₅ requires C, 58.72; H, 4.53; N, 5.48. found C, 58.70;H, 4.24; N, 5.25

Synthesis of3-(4-bromophenyl)-3-((1-(hydroxymethyl)cyclopropyl)methoxy)-2-(4-nitrobenzyl)isoindolin-1-one(NU8391)

The named compound was synthesised from NU8389 (0.50 g, 1.14 mmol) and1,1-bis(hydroxymethyl)cyclopropane (0.22 mL, 2.28 mmol) using GeneralProcedure C, purification by chromatography (Biotage SP4; 10%-50%EtOAc/petrol) and obtained as white crystals (0.47 g, 78%).

¹H NMR (300 MHz, CDCl₃) δ 0.13-0.22 (m, 2H, CH₂), 0.42-0.43 (m, 2H,CH₂), 2.78-2.85 (m, 2H, OCH₂), 3.44-3.52 (m, 2H, HOCH₂), 4.50 (s, 2H,N—CH₂), 7.09-7.18 (m, 3H, Ar—H), 7.28-7.33 (m, 4H, Ar—H), 7.52-7.55 (m,2H, Ar—H), 7.89-7.93 (m, 1H, C(O)═C═CH), 7.96-8.01 (m, 2H, O₂N—C—CH)

¹³C NMR (CDCl₃, 75 MHz), δ 8.90, 22.64, 42.70, 67.72, 94.95, 123.33,123.48, 123.59, 124.14, 128.59, 130.01, 130.20, 130.51, 131.94, 133.47,138.00, 144.95, 145.42, 147.61, 168.54

IR: 697, 759, 805, 855, 928, 1007, 1065, 1097, 1177, 1275, 1341, 1383,1426, 1466, 1519, 1605, 1685, 2873, 2917, 3078 cm⁻¹

LCMS (DMSO): Rt=3.88 min (on 5 min column)

UV (in EtOH): λ max=267 nm

Rf: 0.32 (50% EtOAc/petrol)

MP: 76-77° C.

CHN: C₂₆H₂₃BrN₂O₅ requires C, 59.67; H, 4.43; N, 5.35. found C, 59.71;H, 4.39; N, 5.17

Synthesis of3-(4-bromophenyl)-3-(3-hydroxypropoxy)-2-(4-nitrobenzyl)isoindolin-1-one(NU8392)

The named compound was synthesised from NU8389 (0.50 g, 1.14 mmol) and1,3-propanediol (0.16 mL, 2.28 mmol) using General Procedure C,purification by chromatography (Biotage SP4; 10%-50% EtOAc/petrol) andobtained as a pale yellow oil (0.40 g, 70%).

¹H NMR (300 MHz, CDCl₃) δ 1.46-1.61 (m, 2H, OCH₂—CH₂), 2.92 (td, J=6.0,1.9

Hz, O—CH₂), 3.60 (t, J=6.1 Hz, HO—CH₂), 4.34 and 4.60 (dd: AB, J=15.1Hz, 2H, N—CH₂), 6.95-6.99 (m, 1H, Ar—H), 7.10-7.17 (m, 2H, Ar—H),7.27-7.39 (m, 4H, Ar—H), 7.50-7.55 (m, 2H, Ar—H), 7.89-7.92 (m, 1H,C(O)═C═CH), 7.97-8.04 (m, 2H, O₂N—C—CH)

¹³C NMR (CDCl₃, 75 MHz), δ 32.45, 42.71, 60.23, 60.80, 95.21, 123.31,123.48, 123.61, 124.17, 128.57, 130.24, 130.47, 131.75, 131.99, 133.40,137.99, 145.07, 145.43, 147.64, 168.58

IR: 697, 760, 805, 853, 927, 1010, 1063, 1098, 1177, 1278, 1339, 1381,1423, 1467, 1518, 1603, 1688, 2879, 2925, 3409 cm⁻¹

LCMS (DMSO): Rt=3.73 min (on 5 min column)

UV (in EtOH): λ max=266 nm

Rf: 0.24 (50% EtOAc/petrol).

CHN: C₂₄H₂₁BrN₂O₅ requires C, 57.96; H, 4.26; N, 5.63. found C, 58.16;H, 4.28; N, 5.43

Synthesis of3-(4-chlorophenyl)-3-hydroxy-4-methyl-2-(4-nitrobenzyl)isoindolin-1-one(NU8393) and3-(4-chlorophenyl)-3-hydroxy-7-methyl-2-(4-nitrobenzyl)isoindolin-1-one(NU8394)

The named compounds were synthesised from a mixture of2-(4-chlorobenzoyl)-3-methylbenzoic acid and2-(4-chlorobenzoyl)-6-methylbenzoic acid (2 g, 7.28 mmol) and4-nitrobenzylamine hydrochloride (1.51 g, 8.01 mmol) using GeneralProcedure B, purified by chromatography (Biotage SP4; 10%-20%EtOAc/petrol) and obtained as a yellow solid (NU8393) and cream solid(NU8394) (1.45 g, 49%, ratio of 4- and 7-isomers is 20:1).

Analysis of Major Isomer (NU8393):

¹H NMR (300 MHz, CDCl₃) δ 2.07 (s, 3H, —CH₃), 4.02 (br s, 1H, OH), 4.16and 4.46 (dd, J=15.3 Hz, 2H, N—CH₂—), 7.19-7.23 (m, 6H, Ar—H), 7.29-7.31(dd, J=7.5, 1.5 Hz, 1H, —CH—CH—C(Me), 7.39-7.44 (t, J=7.6 Hz, 1H,—CH—CH—C(Me)-), 7.60-7.62 (dd, J=7.3, 1.1 Hz, 1H, C(O)—C═CH—), 7.93-7.97(m, 2H, —CH—NO₂)

¹³C NMR (CDCl₃, 75 MHz), δ 19.86, 42.76, 90.65, 121.58, 123.67, 125.66,126.70, 128.39, 129.02, 129.69, 131.33, 132.01, 134.21, 136.85, 139.43,145.52, 148.29, 166.89

IR: 695, 729, 760, 800, 851, 932, 982, 1014, 1075, 1193, 1271, 1341,1397, 1487, 1517, 1609, 1680, 3238 cm⁻¹

LCMS (DMSO): Rt=3.75 min (on 5 min column)

HPLC purity (as area %): >95

UV (in EtOH): λ max=269 nm

EI-MS: calculated mass of ion 409.0950 [M+H]⁺, measured mass of ion409.0951 [M+H]⁺

Rf: 0.47 (50% EtOAc/petrol)

MP: 175-176° C.

CHN: C₂₂H_(N)ClN₂O₄ requires C, 64.63; H, 4.19; N, 6.85. found C, 64.78;H, 4.40; N, 6.58

Analysis of Minor Isomer (NU8394):

¹H NMR (300 MHz, CDCl₃) δ 2.62 (s, 3H, —CH₃), 3.79 (br s, 1H, OH), 4.30and 4.61 (dd, J=15.3 Hz, 2H, N—CH₂—), 7.07-7.19 (m, 1H, Ar—H), 7.21-7.26(m, 5H, Ar—H), 7.31-7.41 (m, 3H, Ar—H), 7.97-8.00 (m, 2H, —CH—NO₂)

¹³C NMR (CDCl₃, 75 MHz), δ 17.43, 42.78, 90.68, 120.41, 123.74, 126.76,127.30, 128.18, 129.13, 129.79, 132.57, 133.06, 135.31, 137.40, 138.59,145.87, 149.32, 168.65

IR: 696, 725, 779, 802, 854, 932, 966, 1011, 1088, 1171, 1199, 1279,1337, 1378, 1423, 1481, 1516, 1602, 1678, 2852, 2923, 3338 cm⁻¹

LCMS (DMSO): Rt=3.70 min (on 5 min column)

HPLC purity (as area %): 90

UV (in EtOH): λ max=270 nm

EI-MS: calculated mass of ion 409.0950 [M+H]⁺, measured mass of ion409.0945 [M+H]⁺

Rf: 0.58 (50% EtOAc/petrol)

MP: 158-159° C.

Synthesis of3-(4-chloro-phenyl)-3-(1-hydroxymethyl-cyclopropylmethoxy)-2-(4-nitro-benzyl)-2,3-dihydro-isoindol-1-one(NU8354)

The named compound was synthesised from NU8260 (400 mg, 1.01 mmol) and1,1-bis(hydroxymethyl)cyclopropane (0.19 mL, 2.03 mmol) using GeneralProcedure C, purified by chromatography (Biotage SP4; 10%-50%EtOAc/petrol) and obtained as a cream solid (442 mg, 92%).

¹H NMR (300 MHz, CDCl₃) δ 0.12-0.22 (m, 2H, CH₂), 0.40-0.43 (m, 2H,CH₂), 2.81 (s, 2H, OCH₂), 3.43-3.51 (m, 2H, HOCH₂), 4.49 (s, 2H, N—CH₂),7.12-7.19 (m, 5H, Ar—H), 7.29-7.32 (m, 2H, Ar—H), 7.52-7.55 (m, 2H,Ar—H), 7.89-7.92 (m, 1H, C(O)═C═CH), 7.98-8.01 (m, 2H, O₂N—C—CH)

¹³C NMR (CDCl₃, 75 MHz), δ 8.90, 8.94, 22.68, 42.75, 67.84, 94.96,123.48, 123.60, 124.16, 128.30, 128.98, 130.21, 130.50, 131.87, 133.45,135.29, 137.47, 144.95, 145.52, 168.54

IR: 702, 769, 820, 858, 928, 955, 1009, 1036, 1065, 1090, 1176, 1231,1277, 1341, 1383, 1427, 1470, 1514, 1599, 1705, 2878, 3076, 3471 cm⁻¹

LCMS (DMSO): Rt=3.78 min (on 5 min column)

UV (in EtOH): λ max=267 nm

EI-MS: calculated mass of ion 478.1290 [M+H]⁺, measured mass of ion478.1291 [M+H]⁺

Rf: 0.32 (50% EtOAc/petrol)

MP: 148-149° C.

CHN: C₂₆H₂₃ClN₂O₅ requires C, 65.21; H, 4.84; N, 5.85. found C, 65.21;H, 4.99; N, 5.58

Separation of enantiomers achieved by chiral preparative HPLC (DaicelChiralpak AD-H 250×10 mm; Hexane/Ethanol (4:1))

NU8354A (Yellow Solid)

Optical rotation: Specific rotation [α]=+22.66° (at 24.8° C.,wavelength=589 nm, tube length=0.25 dm, concentration=0.406 g/100 ml)

NU8354B (Off-White Solid)

Optical Rotation Specific rotation [α]=−20.10° (at 24.8° C.,wavelength=589 nm, tube length=0.25 dm, concentration=0.398 g/100 ml)

Synthesis of3-(4-chlorophenyl)-3-hydroxy-5-methyl-2-(4-nitrobenzyl)isoindolin-1-one(NU8395) and3-(4-chlorophenyl)-3-hydroxy-6-methyl-2-(4-nitrobenzyl)isoindolin-1-one(NU8412)

The named compounds were synthesised from a mixture of2-(4-chlorobenzoyl)-4-methylbenzoic acid and2-(4-chlorobenzoyl)-5-methylbenzoic acid (2 g, 7.28 mmol) and4-nitrobenzylamine hydrochloride (1.51 g, 8.01 mmol) using GeneralProcedure B, purified by chromatography (Biotage SP4; 10%-20%EtOAc/petrol) and obtained as a white solid (NU8395) and white solid(NU8412) (1.25 g, 42%, ratio of 5- and 6-isomers is 2:1).

Analysis of Major Isomer (NU8395):

¹H NMR (300 MHz, CDCl₃) δ 1.36 (s, 9H, —C(CH₃)), 3.64 (br s, 1H, OH),4.21 and 4.64 (dd, J=15.3 Hz, 2H, N—CH₂—), 7.19-7.24 (dd, J=1.3 Hz 1H,—C—CH═C(Me)), 7.21-7.22 (m, 4H, Ar—H), 7.25-7.34 (m, 3H, Ar—H),7.65-7.68 (dd, J=7.7 Hz, 1H, C(O)—C═CH—), 7.97-8.01 (m, 2H, —CH—NO₂)

¹³C NMR (CDCl₃, 75 MHz), δ 22.12, 40.91, 94.13, 123.73, 124.04, 128.17,129.16, 129.76, 130.42, 132.88, 134.89, 136.93, 139.91, 140.77, 141.02,145.33, 148.58, 166.11

IR: 696, 729, 745, 770, 788, 800, 833, 853, 929, 961, 988, 1015, 1044,1089, 1103, 1128, 1155, 1184, 1203, 1271, 1309, 1341, 1390, 1427, 1491,1516, 1603, 1623, 1660, 1937, 2202, 2851, 2918, 3053, 3088, 3231 cm⁻¹

LCMS (DMSO): Rt=3.74 min (on 5 min column)

HPLC purity (as area %): >98

UV (in EtOH): λ max=268 nm

EI-MS: calculated mass of ion 426.1215 [M+NH₄]⁺, measured mass of ion426.1212 [M+NH₄]⁺

Rf: 0.50 (50% EtOAc/petrol)

MP: 221-222° C.

CHN: C₂₂H₁₇ClN₂O₄ requires C, 64.63; H, 4.19; N, 6.85. found C, 64.76;H, 4.13, N: 6.58

Analysis of Minor Isomer (NU8412):

¹H NMR (300 MHz, CDCl₃) δ 2.40 (s, 3H, —CH₃), 4.27 (br s, 1H, OH), 4.26and 4.56 (dd, J=15.3 Hz, 2H, N—CH₂—), 7.13-7.21 (m, 5H, Ar—H), 7.25-7.28(m, 2H, Ar—H), 7.30-7.34 (m, 1H, —CH—CH—C(Me)-), 7.43-7.45 (dd, J=1.2Hz, 1H, C(O)—C═CH—), 7.93-7.96 (m, 2H, —CH—NO₂)

¹³C NMR (CDCl₃, 75 MHz), δ 21.64, 42.74, 91.34, 122.82, 123.65, 124.24,128.18, 129.04, 129.77, 130.52, 134.42, 135.21, 137.27, 140.05, 140.80,145.67, 146.24, 168.25

IR: 704, 731, 770, 787, 806, 831, 854, 932, 1013, 1061, 1090, 1132,1174, 1199, 1256, 1310, 1343, 1383, 1433, 1491, 1518, 1607, 1660, 1678,1983, 2018, 2224, 2255, 2853, 2922, 3320, 3342 cm⁻¹

LCMS (DMSO): Rt=4.09 min (on 5 min column)

UV (in EtOH): λ max=268 nm

EI-MS: calculated mass of ion 407.0804 [M−H]⁻, measured mass of ion407.0800 [M−H]⁻

Rf: 0.53 (50% EtOAc/petrol)

MP: 208-209° C.

CHN: C₂₂H₁₇ClN₂O₄ requires C, 64.63; H, 4.19; N, 6.85. found C, 64.95;H, 4.19; N, 6.83

Synthesis of5-tert-butyl-3-(4-chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl)isoindolin-1-one(NU8396) and6-tert-butyl-3-(4-chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl)isoindolin-1-one(NU8397)

The named compounds were synthesised from a mixture of4-tert-butyl-2-(4-chlorobenzoyl)benzoic acid and5-tert-butyl-2-(4-chlorobenzoyl)benzoic acid (2.31 g, 7.28 mmol) and4-nitrobenzylamine hydrochloride (1.51 g, 8.01 mmol) using GeneralProcedure B, purified by chromatography (Biotage SP4; 10%-20%EtOAc/petrol) and obtained as a white solid (NU8396) and cream solid(NU8397) (2.43 g, 74%, ratio of 5- and 6-isomers is 1.3:1).

Analysis of Major Isomer (NU8396):

¹H NMR (300 MHz, CDCl₃) δ 4.25 and 4.68 (dd, J=15.4 Hz, 2H, N—CH₂—),7.22-7.27 (m, 5H, Ar—H), 7.32-7.35 (m, 2H, Ar—H), 7.54-7.57 (dd, J=8.06,1.62 Hz, 1H, —CH—CH—C(^(t)Bu)-), 7.72-7.75 (dd, J=8.0 Hz, 1H,C(O)—C═CH—CH—C(^(t)Bu)-), 8.00-8.03 (m, 2H, —CH—NO₂)

¹³C NMR (CDCl₃, 75 MHz), δ 31.58, 41.48, 41.54, 91.51, 119.64, 123.70,125.51, 126.81, 127.26, 128.20, 129.13, 129.71, 133.51, 134.89, 138.81,141.91, 145.87, 149.50, 167.81

IR: 701, 802, 850, 932, 958, 1011, 1052, 1090, 1194, 1277, 1339, 1395,1423, 1487, 1515, 1603, 1666, 2160, 2955, 3260, 3461 cm⁻¹

LCMS (DMSO): Rt=3.74 min (on 5 min column)

HPLC purity (as area %): >93

UV (in EtOH): λ max=268 nm

EI-MS: calculated mass of ion 451.1419 [M+H]⁺, measured mass of ion451.1413 [M+H]⁺

Rf: 0.58 (50% EtOAc/petrol)

MP: 247-248° C.

CHN: C₂₅H₂₃ClN₂O₄ requires C, 66.59; H, 5.14; N, 6.21. found C, 66.64;H, 5.19; N, 5.89

Analysis of Minor Isomer (NU8397):

¹H NMR (300 MHz, CDCl₃) δ 1.36 (s, 9H, —C(CH₃)), 3.64 (br s, 1H, OH),4.15 and 4.59 (dd, J=15.3 Hz, 2H, N—CH₂—), 7.19-7.24 (m, 5H, Ar—H),7.27-7.30 (m, 2H, Ar—H), 7.57-7.61 (dd, J=8.04, 1.81 Hz, 1H,—CH—CH—C(^(t)Bu)-), 7.90-7.91 (dd, J=1.5 Hz, 1H, C(O)—C═CH—C(^(t)Bu)-),7.97-8.00 (m, 2H, —CH—NO₂)

¹³C NMR (CDCl₃, 75 MHz), δ 31.63, 35.54, 42.75, 91.31, 120.95, 122.56,123.67, 128.18, 129.08, 129.75, 130.93, 145.75

IR: 706, 727, 808, 833, 849, 901, 924, 939, 1013, 1049, 1088, 1134,1198, 1258, 1271, 1309, 1345, 1393, 1439, 1487, 1517, 1603, 1682, 1792,1898, 1923, 1948, 1969, 2065, 2104, 2217, 2365, 2866, 2963, 3381 cm⁻¹

LCMS (DMSO): Rt=3.96 min (on 5 min column)

HPLC purity (as area %): >97

UV (in EtOH): λ max=268 nm

EI-MS: calculated mass of ion 451.1419 [M+H]⁺, measured mass of ion451.1420 [M+H]⁺

Rf: 0.64 (50% EtOAc/petrol)

MP: 226-227° C.

CHN: C₂₅H₂₃ClN₂O₄+0.5EtOAc requires C, 65.52; H, 5.50; N, 5.66. found C,65.44; H, 5.27; N, 5.47

Synthesis of4-chloro-3-(4-chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl)isoindolin-1-one(NU8398)

The named compound was synthesised from a mixture of3-chloro-2-(4-chlorobenzoyl)benzoic acid and6-chloro-2-(4-chlorobenzoyl)benzoic acid (2.15 g, 7.28 mmol) and4-nitrobenzylamine hydrochloride (1.51 g, 8.01 mmol) using GeneralProcedure B, purified by chromatography (Biotage SP4; 10%-20%EtOAc/petrol) and obtained as a white solid (1.96 g, 63%).

¹H NMR (300 MHz, CDCl₃) δ 4.28 (br s, 1H, OH), 4.28 and 4.63 (dd, AB,J=15.4 Hz, N—CH₂—), 7.19-7.22 (m, 4H, Ar—H), 7.31-7.33 (m, 2H, Ar—H),7.44-7.49 (m, 2H, Ar—H), 7.72-7.75 (dd, J=3.2, 8.5 Hz, 1H, (C(O)—C═CH—),7.98-8.00 (m, 2H, —CH—NO₂)

¹³C NMR (CDCl₃, 75 MHz), δ 42.37, 90.90, 122.20, 123.22, 123.35, 128.21,128.68, 129.44, 129.75, 131.69, 131.89, 134.14, 135.15, 147.37, 163.60,163.86

IR: 696, 729, 759, 808, 856, 932, 996, 1070, 1092, 1144, 1174, 1271,1342, 1397, 1462, 1518, 1592, 1682, 2026, 2171, 3220 cm⁻¹

LCMS (DMSO): 8.47 min (on 12 min column)

UV (in EtOH): λ max=268 nm

EI-MS: calculated mass of ion 429.0403 [M+H]⁺, measured mass of ion429.0401 [M+H]⁺

Rf: 0.47 (50% EtOAc/petrol)

MP: 202-203° C.

CHN: C₂₁H₁₄Cl₂N₂O₄+0.2EtOAc requires C, 58.59; H, 3.52; N, 6.27. foundC, 58.27; H, 3.21; N, 6.48

Synthesis of6-tert-butyl-3-(4-chlorophenyl)-3-((1-(hydroxymethyl)cyclopropyl)methoxy)-2-(4-nitrobenzyl)isoindolin-1-one(NU8399)

The named compound was synthesised from NU8397 (200 mg, 0.44 mmol) and1,1-bis(hydroxymethyl)cyclopropane (0.09 mL, 0.89 mmol) using GeneralProcedure C, purified by chromatography (Biotage SP4; 10%-50%EtOAc/petrol) and obtained as white crystals (182 mg, 75%).

¹H NMR (300 MHz, CDCl₃) δ 0.20-0.30 (m, 2H, cyclopropane CH₂), 0.47-0.50(m, 2H, Cyclopropane CH₂), 1.42 (s, 9H, C(CH₃)), 2.12 (br s, 1H, OH),2.86-2.90 (dd, AB, J=9.5 Hz, C—O—CH₂—), 3.51-3.60 (m, 2H, CH₂OH), 4.57(s, 2H, N—CH₂—), 7.13-7.16 (dd, J=7.9 Hz, 1H, CH—CH—C(^(t)Bu)),7.21-7.26 (m, 4H, Ar—H), 7.37-7.40 (m, 2H, Ar—H), 7.62-7.65 (dd, J=8.0,1.8 Hz, 1H, —CH—CH—C(^(t)Bu)), 7.99-8.00 (dd, J=1.3 Hz, 1H—C(O)—C—CH—),8.04-8.06 (m, 2H, —CH—NO₂)

¹³C NMR (CDCl₃, 75 MHz), δ 6.48, 20.00, 31.73, 41.69, 42.50, 68.00,68.14, 94.86, 120.99, 122.07, 123.01, 123.62, 126.57, 126.82, 128.29,128.95, 130.17, 135.15, 137.97, 139.28, 145.28, 147. 59, 167.20

IR: 649, 704, 729, 754, 801, 813, 835, 853, 927, 949, 1012, 1031, 1059,1091, 1135, 1177, 1200, 1259, 1280, 1313, 1342, 1377, 1398, 1434, 1463,1489, 1520, 1600, 1686, 2013, 2093, 2139, 2165, 2189, 2208, 2870, 2922,2960, 3001, 3075, 3428 cm⁻¹

LCMS (DMSO): Rt=3.72 min (on 5 min column)

HPLC purity (as area %): >98

UV (in EtOH): λ max=267 nm

EI-MS: calculated mass of ion 552.2260 [M+NH₄]⁺, measured mass of ion552.2253 [M+NH₄]⁺

Rf: 0.53 (50% EtOAc/petrol)

MP: 104-105° C.

CHN: C₃₀H₃₁ClN₂O₅ requires C, 67.35; H, 5.84; N, 5.24. found C, 67.26;H, 5.89; N, 5.21

Synthesis of5-tert-butyl-3-(4-chlorophenyl)-3-((1-(hydroxymethyl)cyclopropyl)methoxy)-2-(4-nitrobenzyl)isoindolin-1-one(NU8400)

The named compound was synthesised from NU8396 (200 mg, 0.44 mmol) and1,1-bis(hydroxymethyl)cyclopropane (0.09 mL, 0.89 mmol) using GeneralProcedure C, purified by chromatography (Biotage SP4; 10%-50%EtOAc/petrol) and obtained as white crystals (43 mg, 18%).

¹H NMR (300 MHz, CDCl₃) δ 0.10-0.13 (m, 2H, cyclopropane CH₂), 0.41-0.44(m, 2H, Cyclopropane CH₂), 1.26 (s, 9H, C(CH₃)), 1.69 (br s, 1H, OH),2.71-2.78 (m, 2H, C—O—CH₂—), 3.41-3.52 (dd, AB, J=11.0 Hz, CH₂OH),4.38-4.53 (dd, J=15.2 Hz, 2H, N—CH₂—), 7.13-7.15 (dd, J=1.1 Hz, 1H,C—CH—C(^(t)Bu)), 7.16-7.21 (m, 4H, Ar—H), 7.29-7.32 (m, 2H, Ar—H),7.55-7.58 (dd, J=8.1, 1.6 Hz, 1H, —CH—CH—C(^(t)Bu)), 7.81-7.83 (dd,J=7.9 Hz, 1H—C(O)—C—CH—), 7.98-8.01 (m, 2H, —CH—N₂)

¹³C NMR (CDCl₃, 75 MHz), δ 8.95, 9.02, 22.69, 31.56, 42.72, 42.80,67.94, 67.99, 93.23, 123.55, 123.70, 126.59, 127.79, 128.27, 128.91,130.03, 130.11, 131.28, 133.01, 135.15, 138.50, 145.17, 147.27, 168.57

IR: 663, 691, 704, 730, 801, 843, 872, 893, 934, 951, 1013, 1059, 1091,1132, 1189, 1240, 1260, 1281, 1342, 1382, 1398, 1425, 1465, 1490, 1520,1605, 1686, 2010, 2872, 2926, 3073, 3413 cm⁻¹

LCMS: Rt=3.80 min (on 5 min column)

HPLC purity (as area %)=>97

UV (in EtOH): λ max=268 nm

EI-MS: calculated mass of ion 552.2260 [M+NH₄]⁺, measured mass of ion552.2260 [M+NH₄]⁺

Rf: 0.50 (50% EtOAc/petrol)

MP: 89-90° C.

CHN: C₃₀H₃₁ClN₂O₅ requires C, 67.35; H, 5.84; N, 5.24. found C, 67.45;H, 6.01; N, 5.05

Synthesis of3-(4-chlorophenyl)-3-((1-(hydroxymethyl)cyclopropyl)methoxy)-5-methyl-2-(4-nitrobenzyl)isoindolin-1-one(NU8401)

The named compound was synthesised from a mixture of NU8395 and NU8412(200 mg, 0.49 mmol) and 1,1-bis(hydroxymethyl)cyclopropane (0.10 mL,0.98 mmol) using General Procedure C, purified by chromatography(Biotage SP4; 10%-30% EtOAc/petrol) and obtained as a white solid (116mg, 48%).

¹H NMR (300 MHz, CDCl₃) δ 0.07-0.28 (m, 2H, cyclopropane CH₂), 0.34-0.57(m, 2H, cyclopropane CH₂), 2.33-2.40 (s, 3H, CH₃), 2.74-2.88 (m, 2H,C—O—CH₂—), 3.39-3.56 (m, 2H, CH₂OH)), 4.35-4.62 (s, 2H, N—CH₂—),6.90-6.96 (m, 1H, Ar—H), 7.11-7.20 (m, 4H, Ar—H), 7.27-7.41 (m, 3H,Ar—H), 7.74-7.84 (dd, J=7.8 Hz, 1H, —C(O)—C═CH—), 7.92-8.08 (m, 2H,—CH—NO₂)

¹³C NMR (CDCl₃, 75 MHz), δ 8.89, 8.98, 22.16, 22.70, 42.71, 67.87,67.98, 94.77, 123.59, 123.87, 124.02, 128.22, 128.29, 128.96, 130.20,131.51, 135.10, 135.26, 137.63, 140.40, 145.85, 147.74, 166.79

IR: 662, 683, 704, 734, 766, 781, 801, 829, 845, 860, 893, 916, 937,961, 1011, 1032, 1077, 1094, 1134, 1156, 1175, 1211, 1273, 1342, 1391,1420, 1463, 1487, 1518, 1609, 1682, 1925, 2162, 2853, 2882, 2925, 3009,3086, 3406 cm⁻¹

LCMS (DMSO): Rt=3.73 min (on 5 min column)

HPLC purity (as area %): >96

UV (in EtOH): λ max=266 nm

EI-MS: calculated mass of ion 493.1525 [M+H]⁺, measured mass of ion493.1521 [M+H]⁺

Rf=0.53 (50% EtOAc/petrol)

MP: 177-178° C.

CHN: C₂₇H₂₅ClN₂O₅ requires C, 65.79; H, 5.11; N, 5.68. found C, 65.57;H, 5.15; N, 5.45

Synthesis of3-(4-chlorophenyl)-3-((1-(hydroxymethyl)cyclopropyl)methoxy)-4-methyl-2-(4-nitrobenzyl)isoindolin-1-one(NU8405)

The named compound was synthesised from NU8393 (413 mg, 1.01 mmol) and1,1-bis(hydroxymethyl)cyclopropane (0.19 mL, 2.03 mmol) using GeneralProcedure C, purified by chromatography (Biotage SP4; 20%-40%EtOAc/petrol) and obtained as a yellow solid (252 mg, 50%).

¹H NMR (300 MHz, CDCl₃) δ −0.07-0.17 (m, 2H, cyclopropane CH₂),0.25-0.34 (m, 2H, Cyclopropane CH₂), 1.81 (s, 3H, CH₃), 1.92 (br s, 1H,OH), 2.60-2.83 (dd, AB, J=9.4 Hz, C—O—CH₂—), 3.33-3.41 (m, 2H, CH₂OH),4.19 and 4.42 (dd, AB, J=15.3 Hz, 2H, N—CH₂—), 6.88-6.97 (m, 2H, Ar—H)),7.04-7.06 (m, 2H, Ar—H), 7.11-7.13 (dd, J=7.5 Hz, 1H, —CH—C(Me)),7.26-7.30 (t, J=7.5 Hz, 1H, —CH—CH—C(Me)), 7.57-7.60 (dd, J=7.4 Hz,1H—C(O)—C—CH—), 7.75-7.79 (m, 2H, —CH—N₂)

¹³C NMR (CDCl₃, 75 MHz), δ 8.82, 8.95, 17.30, 17.40, 22.68, 42.28,67.60, 94.73, 121.62, 123.47, 128.45, 128.82, 129.94, 130.81, 132.18,134.69, 135.24, 136.92, 142.26, 145.11, 147.50, 168.60

IR: 696, 762, 797, 930, 1013, 1067, 1186, 1229, 1286, 1340, 1393, 1427,1487, 1520, 1605, 1677, 2864, 3455 cm⁻¹

LCMS (DMSO): Rt=3.68 min (on 5 min column)

HPLC purity (as area %): >96

UV (in EtOH): λ max=268 nm

EI-MS: calculated mass of ion 493.1525 [M+H]⁺, measured mass of ion493.1523 [M+H]⁺

Rf: 0.36 (50% EtOAc/petrol); MP: 146-147° C.

CHN: C₂₇H₂₅ClN₂O₅ requires C, 65.79; H, 5.11; N, 5.68. found C, 65.78;H, 4.81; N, 5.65

Synthesis of4-chloro-3-(4-chlorophenyl)-3-((1-(hydroxymethyl)cyclopropyl)methoxy)-2-(4-nitrobenzyl)isoindolin-1-one(NU8406)

The named compound was synthesised from NU8398 (433 mg, 1.01 mmol) and1,1-bis(hydroxymethyl)cyclopropane (0.19 mL, 2.03 mmol) using GeneralProcedure C, purified by chromatography (Biotage SP4; 20%-50%EtOAc/petrol) and obtained as yellow crystals (321 mg, 62%).

¹H NMR (300 MHz, CDCl₃) δ 0.21-0.42 (m, 2H, cyclopropane CH₂), 0.47-0.54(m, 2H, cyclopropane CH₂), 2.12 (br s, 1H, OH), 2.89-3.05 (m, 2H,C—O—CH₂—), 3.52-3.61 (m, 2H, CH₂OH), 4.30-4.59 (dd, AB, J=15.2 Hz,N—CH₂—), 7.15-7.18 (m, 4H, Ar—H), 7.28-7.33 (m, 2H, Ar—H), 7.48-7.58 (m,2H, Ar—H), 7.87-7.89 (dd, J=7.1, 1.1 Hz, 1H, —C(O)—C═CH—), 7.98-8.01 (m,2H, —CH—NO₂)

¹³C NMR (CDCl₃, 75 MHz), δ 8.84, 8.90, 22.59, 42.54, 67.58, 68.10,94.71, 122.63, 123.56, 128.65, 128.77, 130.09, 130.40, 132.28, 134.37,135.27, 135.55, 135.57, 141.10, 144.62, 147.64, 167.04

IR: 696, 759, 816, 853, 930, 1011, 1074, 1144, 1171, 1234, 1341, 1384,1428, 1462, 1489, 1519, 1699, 2872, 2923, 3422 cm⁻¹

HPLC purity (as area %): >92

UV (in EtOH): λ max=267 nm

EI-MS: calculated mass of ion 530.1244 [M+NH₄]⁺, measured mass of ion530.1242 [M+NH₄]⁺

Rf: 0.30 (50% EtOAc/petrol); MP: 76-77° C.

CHN: C₂₆H₂₂Cl₂N₂O₅ requires C, 60.83; H, 4.32; N, 5.46. found C, 60.68;H, 4.30; N, 5.40

Synthesis of5-bromo-3-(4-chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl)isoindolin-1-one(NU8414) and6-bromo-3-(4-chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl)isoindolin-1-one(NU8413)

The named compounds were synthesised from a mixture of4-bromo-2-(4-chlorobenzoyl)benzoic acid and5-bromo-2-(4-chlorobenzoyl)benzoic acid (2.48 g, 7.28 mmol) and4-nitrobenzylamine hydrochloride (1.51 g, 8.01 mmol) using GeneralProcedure B, purified by chromatography (Biotage SP4; 20% EtOAc/petrol)and obtained as a cream solid (NU8414) and cream solid (NU8413) (1.83 g,53%, ratio of 5- and 6-isomers is −1:1).

Analysis of NU8414:

¹H NMR (300 MHz, MeOD) δ 4.50 and 4.68 (dd, J=15.7 Hz, 2H, N—CH₂—), 4.88(br s, 1H, OH), 7.24-7.32 (m, 4H, Ar—H), 7.37-7.47 (m, 3H, Ar—H),7.73-7.77 (dd, J=9.6 Hz, 1H, —CH—CH—C(Br)-), 7.99-8.01 (m, 1H,C(OH)—C═CH—CBr—), 8.04-8.09 (m, 2H, —CH—NO₂)

¹³C NMR (MeOD, 75 MHz), δ 41.27, 93.73, 123.41, 124.51, 125.12, 127.77,129.08, 129.64, 130.09, 130.90, 133.50, 134.67, 134.72, 140.45, 140.72,142.17, 168.08

IR: 698, 781, 801, 827, 851, 895, 934, 1013, 1043, 1071, 1094, 1128,1253, 1342, 1392, 1415, 1516, 1603, 1676, 2405, 2932, 3227 cm⁻¹

LCMS (DMSO): Rt=4.13 min (on 5 min column)

HPLC purity (as area %)=99

UV (in EtOH): λ max=267 nm

EI-MS: calculated mass of ion 471.9820 [M−H], measured mass of ion471.9818 [M−H]

Rf: 0.59 (50% EtOAc/petrol); MP: 221-222° C.

CHN: C₂H₁₄BrClN₂O₄ requires C, 53.25; H, 2.98; N, 5.91. found C, 53.48;H, 2.99; N, 5.79

Analysis of NU8413:

¹H NMR (300 MHz, CDCl₃) δ 3.94 (br s, 1H, OH), 4.22 and 4.53 (dd, J=15.3Hz, 2H, N—CH₂—), 7.06-7.09 (d, J=8.0 Hz, 1H, CH—CH—C(Br)-), 7.11-7.14(m, 4H, Ar—H), 7.17-7.22 (m, 2H, Ar—H), 7.56-7.60 (dd, J=8.0, 1.8 Hz,1H, —CH—CH—C(Br)-), 7.71-7.72 (dd, J=1.6 Hz, 1H, C(O)—C═CH—), 7.88-7.91(m, 2H, —CH—N₂)

¹³C NMR (CDCl₃, 75 MHz), δ 41.63, 94.94, 122.73, 123.76, 124.66, 127.26,128.07, 129.28, 129.82, 132.04, 133.88, 135.53, 136.67, 143.14, 145.09,145.38, 165.32

IR: 718, 754, 801, 827, 851, 930, 1011, 1063, 1090, 1183, 1273, 1311,1342, 1389, 1437, 1487, 1516, 1599, 1672, 2853, 2923, 3163 cm⁻¹

LCMS (DMSO): Rt=4.21 min (on 5 min column)

HPLC purity (as area %)=98

UV (in EtOH): λ max=267 nm

EI-MS: calculated mass of ion 471.9820 [M-1-1], measured mass of ion471.9814 [M−H]

Rf: 0.66 (50% EtOAc/petrol); MP: 218-219° C.

Synthesis of4-((1-(4-chlorophenyl)-1-((1-(hydroxymethyl)cyclopropyl)methoxy)-3-oxoisoindolin-2-yl)methyl)benzonitrile(NU8415)

The named compound was synthesised from NU8306 (379 mg, 1.01 mmol) and1,1-bis(hydroxymethyl)cyclopropane (0.19 mL, 2.03 mmol) using GeneralProcedure C, purified by chromatography (Biotage SP4; 20%-50%EtOAc/petrol) and obtained as white crystals (221 mg, 51%).

¹H NMR (300 MHz, CDCl₃) δ 0.01-0.19 (m, 2H, cyclopropane CH₂), 0.34-0.42(m, 2H, cyclopropane CH₂), 1.96 (s, 1H, OH), 2.72-2.79 (m, 2H, C—O—CH₂),3.39-3.49 (dd, J=11.3, 18.2 Hz, 2H, CH₂OH)), 4.40 (s, 2H, N—CH₂—),7.05-7.12 (m, 5H, Ar—H), 7.18-7.21 (d, AB quartet, J=8.2 Hz, 2H,—CH—CH═C—CN), 7.35-7.37 (d, AB quartet, J=8.2 Hz, 2H, —CH═C—CN),7.45-7.50 (m, 2H, Ar—H), 7.82-7.85 (m, 1H, —C(O)—C═CH—)

¹³C NMR (CDCl₃, 75 MHz), δ 8.86, 22.61, 42.97, 67.43, 67.60, 94.88,111.53, 118.74, 123.50, 124.06, 128.32, 128.88, 130.09, 130.45, 131.81,132.17, 133.44, 135.07, 137.52, 143.10, 145.51, 168.57

IR: 700, 762, 810, 849, 926, 955, 1009, 1041, 1063, 1276, 1302, 1350,1395, 1425, 1468, 1491, 1605, 1680, 2228, 2854, 2926, 3408 cm⁻¹

LCMS (DMSO): Rt=3.90 min (on 5 min column)

HPLC purity (as area %): >98

UV (in EtOH): λ max=225 nm

EI-MS: calculated mass of ion 459.1470 [M+H]⁺, measured mass of ion459.1471 [M+H]⁺

Rf: 0.32 (50% EtOAc/petrol); MP: 136-137° C.

CHN: C₂₇H₂₃ClN₂O₃ requires C, 70.66; H, 5.05; N, 6.10. found C, 70.39;H, 5.05; N, 6.09

Synthesis of2-(4-chlorobenzyl)-3-(4-chlorophenyl)-3-((1-(hydroxymethyl)cyclopropyl)methoxy)isoindolin-1-one(NU8416)

The named compound was synthesised from NU8314 (200 mg, 0.52 mmol) and1,1-bis(hydroxymethyl)cyclopropane (0.10 mL, 1.04 mmol) using GeneralProcedure C, purified by chromatography (Biotage SP4; 20%-40%EtOAc/petrol) and obtained as white crystals (192 mg, 79%).

¹H NMR (300 MHz, CDCl₃) δ 0.05-0.12 (m, 2H, cyclopropane CH₂), 0.34-0.39(m, 2H, cyclopropane CH₂), 1.97 (s, 1H, OH), 2.63-2.79 (dd, AB, J=9.4Hz, 2H, —C—O—CH₂), 3.34-3.50 (dd, AB, J=11.3 Hz, 2H, CH₂OH)), 4.16-4.49(dd, AB, J=14.9 Hz, 2H, N—CH₂—), 7.07-7.09 (m, 4H, Ar—H), 7.10-7.14 (m,5H, Ar—H), 7.44-7.47 (m, 2H, Ar—H), 7.83-7.86 (m, 1H, —C(O)—C═CH—)

¹³C NMR (CDCl₃, 75 MHz), δ 8.82, 22.55, 42.76, 60.56, 67.70, 95.09,123.33, 124.00, 128.30, 128.57, 128.90, 130.28, 130.94, 132.00, 133.22,133.58, 134.94, 136.32, 137.66, 145.68, 168.48

IR: 700, 728, 761, 803, 812, 847, 924, 953, 1009, 1037, 1067, 1092,1176, 1232, 1285, 1318, 1355, 1383, 1425, 1470, 1487, 1609, 1098, 2882,2927, 3489 cm⁻¹

LCMS (DMSO): Rt=3.91 min (on 5 min column)

HPLC purity (as area %): >99

UV (in EtOH): λ max=222 nm

EI-MS: calculated mass of ion 468.1128 [M+H]⁺, measured mass of ion468.1128 [M+H]⁺ Rf=0.41 (50% EtOAc/petrol); MP: 118-119° C.

Synthesis of2-(4-bromobenzyl)-3-(4-chlorophenyl)-3-((1-(hydroxymethyl)cyclopropyl)methoxy)isoindolin-1-one(NU8417)

The named compound was synthesised from NU8315 (433 mg, 1.01 mmol) and1,1-bis(hydroxymethyl)cyclopropane (0.19 mL, 2.03 mmol) using GeneralProcedure C, purified by chromatography (Biotage SP4; 20%-40%EtOAc/petrol) and obtained as white crystals (333 mg, 64%).

¹H NMR (300 MHz, DMSO) δ −0.02-0.08 (m, 2H, cyclopropane CH₂), 0.26-0.33(m, 2H, cyclopropane CH₂), 2.59-2.80 (dd, AB, J=9.0 Hz, 2H, —C—O—CH₂),3.16-3.45 (m, 2H, CH₂OH)), 4.21-4.42 (dd, AB, J=15.4 Hz, 2H, N—CH₂—),4.45 (s, 1H, OH), 7.03-7.06 (d, J=8.4 Hz, 2H, Ar—H), 7.19-7.22 (m, 3H,Ar—H), 7.29-7.40 (m, 4H, Ar—H), 7.56-7.65 (m, 2H, Ar—H), 7.83-7.85 (m,1H, —C(O)—C═CH—)

¹³C NMR (DMSO, 75 MHz), δ 7.94, 22.19, 40.98, 65.96, 73.03, 94.24,120.42, 123.40, 123.59, 123.83, 128.45, 128.66, 130.99, 131.16, 131.40,131.76, 133.49, 137.13, 138.13, 143.48, 167.64

IR: 679, 721, 760, 795, 814, 849, 922, 953, 1008, 1036, 1066, 1092,1177, 1232, 1285, 1317, 1356, 1385, 1420, 1471, 1612, 1692, 2586, 2884,2944, 3499 cm⁻¹

LCMS (DMSO): Rt=4.06 min (on 5 min column)

HPLC purity (as area %): >99

UV (in EtOH): λ max=223 nm

EI-MS: calculated mass of ion 512.0623 [M+H]⁺, measured mass of ion512.0627 [M+H]⁺

Rf=0.41 (50% EtOAc/petrol); MP: 161-162° C.

CHN: C₂₆H₂₃BrClNO₃ requires C, 60.89; H, 4.52; N, 2.73. found C, 60.95;H, 4.61; N, 2.74

Synthesis of3-(4-chlorophenyl)-2-((R)-1-(4-chlorophenyl)ethyl)-3-((1-(hydroxymethyl)cyclopropyl)methoxy)isoindolin-1-one(NU8418)

The named compound was synthesised from NU8301 (401 mg, 1.01 mmol) and1,1-bis(hydroxymethyl)cyclopropane (0.19 mL, 2.03 mmol) using GeneralProcedure C, purified by chromatography (Biotage SP4; 20%-40%EtOAc/petrol) and obtained as a pale yellow oil (146 mg, 30%).

¹H NMR (300 MHz, CDCl₃) δ 0.36-0.44 (m, 2H, cyclopropane CH₂), 0.48-0.59(m, 2H, cyclopropane CH₂), 1.88 (d, J=7.3 Hz, 3H, CH₃), 2.30 (br s, 1H,OH), 2.88-3.30 (dd, AB, J=9.5 Hz, 2H, —C—O—CH₂), 3.58-3.67 (m, 2H,CH₂OH)), 4.30-4.38 (q, J=7.2, 14.4 Hz, 1H, N—CH—), 6.96-7.01 (m, 7H,Ar—H), 7.04-7.07 (m, 1H, Ar—H), 7.43-7.47 (m, 2H, Ar—H), 7.80-7.83 (m,1H, —C(O)—C═CH—)

¹³C NMR (CDCl₃, 75 MHz), δ 8.96, 9.03, 20.41, 22.93, 52.80, 60.59,67.83, 95.25, 123.28, 123.75, 128.27, 128.43, 128.68, 129.57, 130.31,133.09, 133.08, 134.82, 137.55, 141.72, 144.99, 168.30

IR: 699, 727, 762, 815, 868, 938, 1011, 1032, 1070, 1088, 1176, 1321,1398, 1467, 1489, 1597, 1682, 2876, 2930, 3389 cm⁻¹

LCMS (DMSO): Rt=4.15 min (on 5 min column)

HPLC purity (as area %): >97

UV (in EtOH): λ max=221 nm

EI-MS: calculated mass of ion 482.1284 [M+H]⁺, measured mass of ion482.1279 [M+H]⁺

Rf=0.42 (50% EtOAc/petrol)

Synthesis of3-(4-chlorophenyl)-2-((S)-1-(4-chlorophenyl)ethyl)-3-((1-(hydroxymethyl)cyclopropyl)methoxy)isoindolin-1-one(NU8419)

The named compound was synthesised from NU8347 (401 mg, 1.01 mmol) and1,1-bis(hydroxymethyl)cyclopropane (0.19 mL, 2.03 mmol) using GeneralProcedure C, purified by chromatography (Biotage SP4; 10%-30%EtOAc/petrol) and obtained as a pale yellow oil (60 mg, 12%).

¹H NMR (300 MHz, CDCl₃) δ 0.36-0.44 (m, 2H, cyclopropane CH₂), 0.48-0.59(m, 2H, cyclopropane CH₂), 1.88 (d, J=7.3 Hz, 3H, CH₃), 2.30 (br s, 1H,OH), 2.88-3.30 (dd, AB, J=9.5 Hz, 2H, —C—O—CH₂), 3.58-3.67 (m, 2H,CH₂OH)), 4.30-4.38 (q, J=7.2, 14.4 Hz, 1H, N—CH—), 6.96-7.01 (m, 7H,Ar—H), 7.04-7.07 (m, 1H, Ar—H), 7.43-7.47 (m, 2H, Ar—H), 7.80-7.83 (m,1H, —C(O)—C═CH—)

¹³C NMR (CDCl₃, 75 MHz), δ 8.96, 9.03, 20.41, 22.93, 52.80, 60.59,67.83, 95.25, 123.28, 123.75, 128.27, 128.43, 128.68, 129.57, 130.31,133.09, 133.08, 134.82, 137.55, 141.72, 144.99, 168.30

IR: 698, 727, 761, 815, 866, 1011, 1031, 1068, 1090, 1176, 1331, 1396,1487, 1583, 1695, 2876, 2923, 3415 cm⁻¹

LCMS (DMSO): Rt=4.19 min (on 5 min column)

HPLC purity (as area %): >98

UV (in EtOH): λ max=221 nm

EI-MS: calculated mass of ion 482.1284 [M+H]⁺, measured mass of ion482.1285 [M+H]⁺

Rf=0.42 (50% EtOAc/petrol)

Synthesis of5-bromo-3-(4-chlorophenyl)-3-((1-(hydroxymethyl)cyclopropyl)methoxy)-2-(4-nitrobenzyl)isoindolin-1-one(NU8424)

The named compound was synthesised from NU8414 (200 mg, 0.42 mmol) and1,1-bis(hydroxymethyl)cyclopropane (0.08 mL, 0.84 mmol) using GeneralProcedure C, purified by chromatography (Biotage SP4; 20%-40%EtOAc/petrol) and obtained as a yellow oil (204 mg, 87%).

¹H NMR (300 MHz, CDCl₃) δ 0.17-0.27 (m, 2H, cyclopropane CH₂), 0.44-0.51(m, 2H, cyclopropane CH₂), 2.05 (s, 1H, OH), 2.83-2.90 (dd, AB, J=9.5Hz, 2H, C—O—CH₂), 3.49-3.58 (dd, AB, J=11.5 Hz, 2H, CH₂OH), 4.45-4.56(dd, AB, J=15.4 Hz, 2H, N—CH₂—), 7.16-7.19 (m, 4H, Ar—H), 7.29-7.33 (m,3H, Ar—H), 7.66-7.70 (dd, J=1.6, 8.0 Hz, 1H, —CH—CH—Br), 7.78-7.80 (dd,J=8.0 Hz, 1H, —C(O)—C═CH—), 7.99-8.02 (d, AB, J=8.7 Hz, 2H, CH—C—NO₂)

¹³C NMR (CDCl₃, 75 MHz), δ 8.90, 22.62, 42.79, 67.47, 67.72, 94.47,123.62, 125.52, 126.92, 128.25, 128.41, 129.12, 130.24, 130.60, 134.00,135.54, 136.72, 144.61, 147.35, 147.71, 167.66

IR: 702, 726, 799, 818, 855, 883, 934, 1011, 1032, 1078, 1128, 1176,1277, 1343, 1387, 1420, 1488, 1521, 1601, 1684, 2854, 2922, 3420 cm⁻¹

HPLC purity (as area %): >86

UV (in EtOH): λ max=267 nm

EI-MS: calculated mass of ion 574.0739 [M+NH₄]⁺, measured mass of ion574.0735 [M+NH₄]⁺

Rf=0.37 (50% EtOAc/petrol); MP: 156-158° C.

Synthesis of6-bromo-3-(4-chlorophenyl)-3-((1-(hydroxymethyl)cyclopropyl)methoxy)-2-(4-nitrobenzyl)isoindolin-1-one(NU8425)

The named compound was synthesised from NU8413 (200 mg, 0.42 mmol) and1,1-bis(hydroxymethyl)cyclopropane (0.08 mL, 0.84 mmol) using GeneralProcedure C, purified by chromatography (Biotage SP4; 20%-40%EtOAc/petrol) and obtained as an orange solid (154 mg, 66%).

¹H NMR (300 MHz, CDCl₃) δ 0.21-0.31 (m, 2H, cyclopropane CH₂), 0.49-0.56(m, 2H, cyclopropane CH₂), 2.13 (br s, 1H, OH), 2.88-2.92 (m, 2H,C—O—CH₂), 3.53-3.62 (dd, AB, J=11.4 Hz, 2H, CH₂OH), 4.52-4.62 (dd, AB,J=15.9 Hz, 2H, N—CH₂—), 7.12-7.14 (d, J=8.0 Hz, 1H, CH—CH—CBr),7.20-7.26 (m, 4H, Ar—H), 7.34-7.38 (m, 2H, Ar—H), 7.72-7.75 (dd, J=1.8,8.0 Hz, 1H, —CH—CH—CBr—), 8.04-8.07 (m, 2H, CH—C—NO₂), 8.09-8.10 (d,J=1.6 Hz, 1H, C(═O)—C═CH—C—Br)

¹³C NMR (CDCl₃, 75 MHz), δ 8.88, 8.93, 22.65, 42.84, 67.50, 67.72,94.75, 123.62, 124.81, 125.13, 127.39, 128.24, 129.07, 130.21, 133.68,133.71, 135.46, 136.52, 136.83, 144.17, 144.60, 147.70, 167.09,

IR: 696, 725, 820, 854, 926, 1011, 1176, 1277, 1341, 1376, 1425, 1489,1519, 1602, 1699, 2924, 3077, 3422 cm⁻¹

HPLC purity (as area %): >98

UV (in EtOH): λ max=267 nm

EI-MS: calculated mass of ion 556.0395 [M]⁺, measured mass of ion556.0389 [M]⁺

Rf=0.41 (50% EtOAc/petrol); MP: 66-68° C.

Synthesis of3-(4-chlorophenyl)-3-(1-(hydroxymethyl)cyclopropyl)methoxy)-2-(pyridin-2-ylmethyl)isoindolin-1-one(NU8429)

The named compound was synthesised from NU8423 (354 mg, 1.01 mmol) and1,1-bis(hydroxymethyl)cyclopropane (0.19 mL, 2.03 mmol) using GeneralProcedure C, purified by chromatography (Biotage SP4; 50%EtOAc/petrol—EtOAc) and obtained as yellow crystals (383 mg, 87%).

¹H NMR (300 MHz, CDCl₃) δ 0.04-0.17 (m, 2H, cyclopropane CH₂), 0.30-0.37(m, 2H, cyclopropane CH₂), 2.57-3.18 (dd, J=9.3 Hz, 2H, C—O—CH₂),3.25-3.63 (dd, J=11.3 Hz, 2H, CH₂OH), 4.10 (br s, 1H OH), 4.36-4.48 (dd,AB, J=15.2 Hz, 2H, N—CH₂—), 6.94-7.02 (m, 3H, ArH), 7.04-7.07 (m, 1H,Ar—H), 7.10-7.15 (m, 2H, Ar—H), 7.19-7.22 (m, H, Ar—H), 7.35-7.42 (m,3H, Ar—H), 7.76-7.79 (m, 1H, —C(O)—C═CH—), 8.22-8.23 (m, 1H, —C═N—CH═)

¹³C NMR (CDCl₃, 75 MHz), δ 8.79, 8.86, 22.64, 45.64, 67.13, 67.55,94.79, 122.33, 123.40, 123.93, 124.07, 128.30, 128.69, 130.18, 132.12,133.09, 134.57, 136.53, 137.86, 145.68, 148.96, 157.14, 168.40

IR: 700, 760, 813, 846, 928, 969, 1009, 1038, 1069, 1088, 1113, 1179,1229, 1279, 1315, 1352, 1377, 1421, 1469, 1595, 1695, 2854, 2909, 3277cm⁻¹

HPLC purity (as area %): >97

UV (in EtOH): λ max=261 nm

EI-MS: calculated mass of ion 435.1470 [M+H]⁺, measured mass of ion435.1471 [M+H]⁺

Rf=0.01 (50% EtOAc/petrol); MP: 123-125° C.

Synthesis of3-(4-chlorophenyl)-5-fluoro-3-hydroxy-2-(4-nitrobenzyl)isoindolin-1-one(NCL-00010485) and3-(4-chlorophenyl)-6-fluoro-3-hydroxy-2-(4-nitrobenzyl)isoindolin-1-one(NCL-00010486)

The named compounds were synthesised from a mixture of2-(4-chlorobenzoyl)-4-fluorobenzoic acid and2-(4-chlorobenzoyl)-5-fluorobenzoic acid (2.39 g, 8.58 mmol) and4-nitrobenzylamine hydrochloride (1.78 g, 9.44 mmol) using GeneralProcedure B, purified by chromatography (Biotage SP4; 25% EtOAc/petrol)and obtained as a cream solid (NCL-00010485) and a yellow solid(NCL-00010486) (2.56 g, 72%, ratio of 5- and 6-isomers is 3:2).

Analysis of Major Isomer (NCL-00010485):

¹H NMR (300 MHz, CDCl₃) δ 4.22-4.66 (dd AB, J=15.4 Hz, 2H, N—CH₂), 5.28(s, 1H, OH), 6.85-6.89 (dd, J=7.7, 2.2 Hz, 1H, C(OH)CCH—C(F)), 7.06-7.13(m, 1H, C(═O)—C═CH—CH═), 7.15-7.22 (m, 4H, —C₆ H ₄Cl), 7.30-7.36 (d AB,J=8.7 Hz, 2H, —C₂ H ₂C₂H₂C(NO₂)), 7.72-7.76 (dd, J=8.3, 4.8 Hz, 1H,C(═O)C═CH—), 7.93-7.96 (d AB, J=8.7 Hz, 2H, —C₂ H ₂C(NO₂))

¹³C NMR (DMSO, 75 MHz), δ 42.35, 86.69, 113.40, 117.58, 123.23, 125.67,126.79, 128.46, 128.74, 129.39, 133.52, 138.52, 140.85, 146.22, 147.54,158.81, 166.24

IR: 710, 775, 804, 832, 933, 970, 1015, 1054, 1092, 1155, 1201, 1263,1342, 1394, 1484, 1519, 1607, 1676, 3235 cm⁻¹

LCMS (DMSO): Rt=4.03 min (on 5 min column)

HPLC purity (as area %): >98

UV (in EtOH): λ max=272 nm

EI-MS: calculated mass of ion 430.0964 [M+NH₄]⁺, measured mass of ion430.0958 [M+NH₄]⁺

Rf=0.50 (50% EtOAc/petrol); MP: 222-224° C.

Analysis of Minor Isomer (NCL-00010486):

¹H NMR (300 MHz, CDCl₃) δ 4.25-4.65 (dd AB, J=15.4 Hz, 2H, N—CH₂), 4.73(s, 1H, OH), 7.13-7.24 (m, 6H, ArH), 7.30-7.33 (d AB, J=8.7 Hz, 2H, —C₂H ₂C₂H₂C(NO₂)), 7.35-7.39 (dd, J=7.2, 2.1 Hz, 1H, C(═O)C═CH—), 7.94-7.98(d AB, J=8.7 Hz, 2H, —C₂ H ₂C(NO₂))

¹³C NMR (DMSO, 75 MHz), δ 42.43, 90.42, 113.29, 116.37, 123.23, 125.58,128.45, 128.70, 129.42, 133.48, 134.84, 138.76, 139.16, 142.38, 146.18,160.19, 166.08

IR: 776, 806, 831, 851, 891, 932, 1013, 1059, 1092, 1173, 1197, 1267,1310, 1342, 1390, 1449, 1484, 1518, 1607, 1680, 2160, 2852, 2925, 3267cm⁻¹

LCMS (DMSO): Rt=4.03 min (on 5 min column)

HPLC purity (as area %): >97

UV (in EtOH): λ max=267 nm

EI-MS: calculated mass of ion 412.0621 [M+H]⁺, measured mass of ion412.0619 [M+H]⁺

Rf=0.58 (50% EtOAc/petrol); MP: 188-190° C.

Synthesis of5,6-dichloro-3-(4-chlorophenyl)-3-(1-(hydroxymethyl)cyclopropyl)methoxy)-2-(4-nitrobenzyl)isoindolin-1-one(NCL-00010487)

The named compound was synthesised from NU8432 (350 mg, 0.75 mmol) and1,1-bis(hydroxymethyl)cyclopropane (0.16 mL, 1.67 mmol) using GeneralProcedure C, purified by chromatography (Biotage SP4; 20%-40%EtOAc/petrol) and obtained as a yellow oil (302 mg, 73%).

¹H NMR (300 MHz, CDCl₃) δ 0.20-0.30 (m, 2H, CH ₂), 0.46-0.50 (m, 2H, CH₂), 2.25 (s, 1H, OH), 2.90 (s, 2H, OCH ₂), 3.50-3.60 (dd AB, J=11.2, 8.8Hz, 2H, HOCH ₂), 4.46-4.58 (dd AB, J=15.2, 5.6 Hz, 2H, N—CH ₂),7.19-7.20 (m, 4H, —C₆ H ₄Cl), 7.29-7.32 (m, 3H, N₂O—C═CH—CH═ andC(O—)C═CH—), 7.98-8.01 (m, 3H, O₂N—C—CH and C(O)═C═CH)

¹³C NMR (CDCl₃, 75 MHz), δ 8.87, 8.90, 22.60, 42.93, 67.27, 67.68,94.36, 123.63, 125.76, 126.01, 128.22, 129.17, 130.23, 131.44, 135.56,135.66, 136.41, 138.31, 144.38, 144.77, 147.72, 166.46

IR: 754, 804, 837, 889, 934, 1011, 1070, 1095, 1166, 1201, 1235, 1339,1402, 1489, 1514, 1601, 1688, 2857, 2923, 3482 cm⁻¹

LCMS (DMSO): Rt=4.65 min (on 5 min column)

HPLC purity (as area %): >99

UV (in EtOH): λ max=266 nm

EI-MS: calculated mass of ion 569.0408 [M+Na]⁺, measured mass of ion569.0408 [M+Na]⁺

Rf=0.45 (50% EtOAc/petrol)

Synthesis of4-((7-chloro-1-(4-chlorophenyl)-1-hydroxy-3-oxoisoindolin-2-yl)methyl)benzonitrile(NCL-00010488) and4-((4-chloro-1-(4-chlorophenyl)-1-hydroxy-3-oxoisoindolin-2-yl)methyl)benzonitrile(73/NCL-00010489)

The named compounds were synthesised from a mixture of3-chloro-2-(4-chlorobenzoyl)benzoic acid and6-chloro-2-(4-chlorobenzoyl)benzoic acid (2.15 g, 7.28 mmol) and4-cyanobenzylamine hydrochloride (1.35 g, 8.01 mmol) using GeneralProcedure B, purified by chromatography (Biotage SP4; 20% EtOAc/petrol)and obtained as a white solid (NCL-00010488) and brown crystals(NCL-00010489) (1.98 g, 66%, ratio of 7- and 4-isomers is 99:1)

Analysis of Major Isomer (NCL-00010488):

¹H NMR (300 MHz, CDCl₃) δ 3.50 (br s, 1H, OH), 4.22-4.61 (dd AB, J=15.3Hz, 2H, N—CH₂—), 7.22-7.30 (m, 6H, ArH), 7.40-7.51 (m, 4H, ArH),7.75-7.78 (dd, J=5.8, 2.6 Hz, 1H, C(O)—C═CH)

¹³C NMR (CDCl₃, 75 MHz), δ 42.97, 91.15, 107.13, 109.45, 122.61, 128.49,129.02, 129.65, 130.01, 130.54, 132.09, 132.13, 132.34, 132.40, 135.02,143.99, 145.12, 165.73

IR: 669, 710, 766, 812, 853, 930, 1002, 1069, 1088, 1146, 1175, 1277,1352, 1404, 1462, 1490, 1582, 1661, 2228, 2919, 3205 cm⁻¹

LCMS (DMSO): Rt=3.99 min (on 5 min column)

HPLC purity (as area %): >97

UV (in EtOH): λ max=227 nm

EI-MS: calculated mass of ion 407.0360 [M−H]⁻, measured mass of ion407.0363 [M−H]⁻

Rf=0.42 (50% EtOAc/petrol); MP: 210-211° C.

Analysis of Minor Isomer (NCL-00010489):

¹H NMR (300 MHz, CDCl₃) δ 4.02 (br s, 1H, OH), 4.29-4.60 (dd AB, J=15.1Hz, 2H, N—CH₂—), 7.16-7.19 (dd, J=7.2, 1.2 Hz, 1H, C(OH)—C═CH—),7.20-7.22 (m, 4H, —C₆ H ₄Cl), 7.26-7.29 (m, 2H, ArH), 7.37-7.47 (m, 4H,ArH)

¹³C NMR (CDCl₃, 75 MHz), δ 43.17, 94.29, 100.01, 115.88, 128.17, 128.53,129.18, 129.92, 131.95, 132.32, 132.77, 134.26, 134.36, 135.53, 136.47,143.28, 144.23, 167.99

IR: 672, 698, 736, 789, 801, 845, 928, 956, 1013, 1088, 1169, 1205,1269, 1350, 1382, 1460, 1489, 1597, 1686, 1790, 2226, 1851, 2922, 3076,3358 cm⁻¹

LCMS (DMSO): Rt=3.95 min (on 5 min column)

HPLC purity (as area %): >88

UV (in EtOH): λ max=227 nm

EI-MS: calculated mass of ion 426.0771 [M+NH₄]⁺, measured mass of ion426.0777 [M+NH₄]⁺

Rf=0.57 (50% EtOAc/petrol); MP: 191-193° C.

Synthesis of2-(4-bromobenzyl)-4-chloro-3-(4-chlorophenyl)-3-hydroxyisoindolin-1-one(NCL-00010490)

The named compound was synthesised from a mixture of3-chloro-2-(4-chlorobenzoyl)benzoic acid and6-chloro-2-(4-chlorobenzoyl)benzoic acid (2.15 g, 7.28 mmol) and4-bromobenzylamine (1.49 g, 8.01 mmol) using General Procedure B,purified by chromatography (Biotage SP4; 20% EtOAc/petrol) and obtainedas a white solid (1.57 g, 47%).

¹H NMR (300 MHz, DMSO) δ 3.40 (br s, 1H, OH), 4.06-4.55 (dd AB, J=15.0Hz, 2H, N—CH₂—), 7.01-7.07 (m, 2H, C—C₂ H ₂C₂H₂Br), 7.23-7.32 (m, 6H,ArH), 7.44-7.50 (m, 2H, ArH), 7.72-7.75 (dd, J=5.7, 2.8 Hz, 1H,C(═O)—C═CH)

¹³C NMR (DMSO, 75 MHz), δ 44.30, 91.18, 121.62, 122.51, 126.33, 127.34,128.29, 128.58, 128.94, 130.81, 131.73, 134.30, 137.00, 139.90, 141.07,145.12, 167.02

IR: 671, 711, 766, 814, 847, 928, 1009, 1072, 1150, 1198, 1285, 1356,1399, 1464, 1489, 1585, 1656, 3193 cm⁻¹

LCMS (DMSO): Rt=4.72 min (on 5 min column)

HPLC purity (as area %): >97

UV (in EtOH): λ max=222 nm

EI-MS: calculated mass of ion 459.9512 [M−H]⁻, measured mass of ion459.9515 [MNH]⁻

Rf=0.57 (50% EtOAc/petrol); MP: 211-212° C.

Synthesis of4-((7-chloro-1-(4-chlorophenyl)-1-((1-(hydroxymethyl)cyclopropyl)methoxy)-3-oxoisoindolin-2-yl)methyl)benzonitrile(NCL-00010492)

The named compound was synthesised from NCL-00010488 (413 mg, 1.01 mmol)and 1,1-bis(hydroxymethyl)cyclopropane (0.19 mL, 2.03 mmol) usingGeneral Procedure C, purified by chromatography (Biotage SP4; 20%-60%EtOAc/petrol) and obtained as white crystals (305 mg, 61%).

¹H NMR (300 MHz, CDCl₃) δ 0.23-0.34 (m, 2H, cyclopropane CH₂), 0.38-0.46(m, 2H, cyclopropane CH₂), 2.28 (s, 3H, OH), 2.78-2.92 (dd AB, J=9.1 Hz,2H, —O—CH₂), 3.43-3.53 (dd AB, J=11.4 Hz, 2H, CH₂OH), 4.30-4.45 (dd AB,J=15.2 Hz, 2H, N—CH₂—), 7.02-7.10 (m, 2H, ArH), 7.12-7.16 (m, 4H, ArH),7.33-7.37 (m, 2H, C₂ H ₂—C(CN)), 7.38-7.42 (m, 1H,—CH—CH═C(Cl)—C—C(OCH₂—), 7.43-7.49 (m, 1H, —CH═C(Cl)—C—C(OCH₂—)),7.78-7.80 (dd, J=7.2, 1.2 Hz, 1H, C(═O)—C═CH)

¹³C NMR (CDCl₃, 75 MHz), δ 8.80, 8.86, 22.54, 42.80, 67.46, 68.04,94.69, 111.59, 118.67, 122.60, 128.66, 128.72, 129.98, 130.36, 132.17,132.25, 134.33, 134.41, 135.17, 135.56, 141.10, 142.72, 167.04

IR: 761, 814, 854, 928, 1012, 1144, 1172, 1233, 1275, 1651, 1384, 1423,1462, 1588, 1697, 2229, 2874, 2920, 3423 cm⁻¹

LCMS (DMSO): Rt=4.02 min (on 5 min column)

HPLC purity (as area %): >96

UV (in EtOH): λ max=226 nm

EI-MS: calculated mass of ion 426.0255 [M+H]⁺, measured mass of ion426.0257 [M+H]⁺

Rf=0.26 (50% EtOAc/petrol); MP: 70-72° C.

Synthesis of2-(4-bromobenzyl)-4-chloro-3-(4-chlorophenyl)-3-((1-(hydroxymethyl)cyclopropyl)methoxy)isoindolin-1-one(NCL-00010493)

The named compound was synthesised from NCL-00010490 (468 mg, 1.01 mmol)and 1,1-bis(hydroxymethyl)cyclopropane (0.19 mL, 2.03 mmol) usingGeneral Procedure C, purified by chromatography (Biotage SP4; 20%-40%EtOAc/petrol) and obtained as white crystals (304 mg, 55%).

¹H NMR (300 MHz, DMSO) δ 0.06-0.27 (m, 2H, cyclopropane CH₂), 0.35-0.40(m, 2H, cyclopropane CH₂), 2.23 (s, 3H, OH), 2.82 (s, 2H, —O—CH₂),3.41-3.50 (dd AB, J=11.8 Hz, 2H, CH₂OH), 4.15-4.38 (dd AB, J=14.9 Hz,2H, N—CH₂—), 6.93-6.96 (d, AB, J=8.4 Hz, 2H, —C₂ H ₂C₂H₂Br), 7.10-7.15(m, 4H, ArH), 7.19-7.23 (d AB, J=8.4 Hz, 2H, C₂ H ₂—C(Br)), 7.37-7.47(m, 2H, ArH), 7.77-7.81 (dd, J=7.1, 1.2 Hz, 1H, C(═O)—C═CH),

¹³C NMR (DMSO, 75 MHz), δ 8.83, 8.97, 22.50, 42.70, 67.65, 68.08, 94.91,121.69, 122.54, 128.72, 130.23, 131.15, 131.59, 132.10, 134.17, 134.63,135.06, 135.63, 136.46, 141.22, 166.96

IR: 713, 759, 818, 924, 951, 1009, 1071, 1144, 1172, 1233, 1349, 1383,1461, 1487, 1587, 1690, 2873, 2923, 3404 cm⁻¹

LCMS (DMSO): Rt=4.27 min (on 5 min column)

HPLC purity (as area %): >94

UV (in EtOH): λ max=222 nm

EI-MS: calculated mass of ion 563.0498 [M+NH₄]⁺, measured mass of ion563.0491 [M+NH₄]⁺

Rf=0.39 (50% EtOAc/petrol); MP: 59-61° C.

Synthesis of3-(4-chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl)-2,3,4,5,6,7-hexahydroisoindol-1-one(NCL-00010494)

The named compound was synthesised from2-(4-chlorobenzoyl)cyclohex-1-enecarboxylic acid (1.00 g, 3.78 mmol) and4-nitrobenzylamine hydrochloride (0.78 g, 4.16 mmol) using GeneralProcedure B, purified by chromatography (Biotage SP4; 30% EtOAc/petrol)and obtained as a yellow solid (0.67 g, 44%).

¹H NMR (300 MHz, CDCl₃) δ 1.60-1.80 (m, 4H,—C(═O)—C—CH₂CH₂CH₂CH₂C—C(OH), 2.20-2.33 (m, 4H, —C(═O)—C—CH ₂CH₂CH₂CH₂C—C(OH)), 4.19-4.61 (dd AB, J=15.6 Hz, 2H, N—CH₂—), 4.73 (s, 1H, OH),7.24-7.30 (m, 4H, —C₆ H ₄Cl), 7.33-7.36 (d AB, J=8.6 Hz, 2H, —C₂ H₂C₂H₂C(NO₂)), 7.97-8.00 (d AB, J=8.6 Hz, 2H, —C₂ H ₂C(NO₂))

¹³C NMR (CDCl₃, 75 MHz), δ 20.31, 21.02, 22.11, 22.26, 42.59, 92.16,123.50, 127.94, 129.09, 129.66, 131.47, 134.97, 136.12, 146.31, 147.42,157.64, 171.18

IR: 697, 727, 797, 824, 853, 914, 968, 1013, 1045, 1092, 1136, 1202,1283, 1339, 1435, 1489, 1520, 1603, 1661, 2853, 2932, 3159 cm⁻¹

LCMS (DMSO): Rt=4.02 min (on 5 min column)

HPLC purity (as area %): >98

UV (in EtOH): λ max=272 nm

EI-MS: calculated mass of ion 399.1106 [M+H]⁺, measured mass of ion399.1112 [M+H]⁺

Rf=0.39 (50% EtOAc/petrol); MP: 144-146° C.

Synthesis of3-(4-chlorophenyl)-5-fluoro-3-((1-(hydroxymethyl)cyclopropyl)methoxy)-2-(4-nitrobenzyl)isoindolin-1-one(NCL-00010495)

The named compound was synthesised from NCL-00010485 (150 mg, 0.36 mmol)and 1,1-bis(hydroxymethyl)cyclopropane (0.07 mL, 0.72 mmol) usingGeneral Procedure C, purified by chromatography (Biotage SP4; 20%-40%EtOAc/petrol) and obtained as white crystals (85 mg, 47%).

¹H NMR (300 MHz, CDCl₃) δ 0.10-0.25 (m, 2H, cyclopropane CH₂), 0.40-0.49(m, 2H, cyclopropane CH₂), 1.69 (s, 3H, OH), 2.84 (s, 2H, —O—CH₂),3.43-3.54 (dd AB, J=11.3 Hz, 2H, CH₂OH), 4.48 (s, 2H, N—CH₂—), 6.82-6.85(dd, J=7.5, 2.0 Hz, 1H, ArH), 7.15-7.17 (m, 4H, —C₆ H ₄Cl), 7.18-7.25(dt, J=8.7, 2.2 Hz, 1H, ArH)), 7.28-7.31 (d AB, J=8.6 Hz, 2H, —C₂ H₂C₂H₂C(NO₂)), 7.87-7.92 (dd, J=8.3, 4.8 Hz, 1H, C(═O)—C═CH), 7.98-8.01(d AB, J=8.7 Hz, 2H, —C₂ H ₂C(NO₂))

¹³C NMR (CDCl₃, 75 MHz), δ 8.91, 22.64, 42.86, 67.62, 67.77, 94.30,110.87, 111.19, 118.11, 118.39, 123.63, 126.40, 128.22, 129.12, 130.22,135.56, 136.88, 144.75, 148.23, 167.48, 168.20

IR: 683, 772, 802, 836, 936, 1013, 1059, 1093, 1150, 1178, 1220, 1264,1342, 1383, 1427, 1487, 1520, 1605, 1697, 2876, 2927, 3082, 3407 cm⁻¹

LCMS (DMSO): Rt=4.12 min (on 5 min column)

HPLC purity (as area %): >98

UV (in EtOH): λ max=267 nm

EI-MS: calculated mass of ion 514.1540 [M+NH₄]⁺, measured mass of ion514.1540 [M+NH₄]⁺

Rf=0.35 (50% EtOAc/petrol); MP: 61-63° C.

Synthesis of3-(4-chlorophenyl)-6-fluoro-3-((1-(hydroxymethyl)cyclopropyl)methoxy)-2-(4-nitrobenzyl)isoindolin-1-one(NCL-00010496)

The named compound was synthesised from NCL-00010486 (150 mg, 0.36 mmol)and 1,1-bis(hydroxymethyl)cyclopropane (0.07 mL, 0.72 mmol) usingGeneral Procedure C, purified by chromatography (Biotage SP4; 20%-40%EtOAc/petrol) and obtained as white crystals (115 mg, 64%).

¹H NMR (300 MHz, CDCl₃) δ 0.22-0.31 (m, 2H, cyclopropane CH₂), 0.48-0.55(m, 2H, cyclopropane CH₂), 2.10 (s, 3H, OH), 2.90 (s, 2H, —O—CH₂),3.52-3.62 (dd AB, J=11.4 Hz, 2H, CH₂OH), 4.51-4.62 (dd AB, J=15.5 Hz,2H, N—CH₂—), 7.19-7.25 (m, 5H, —C₆ H ₄Cl and C(F)═CH—CH═), 7.27-7.30(dd, J=8.6, 2.2 Hz, 1H, —C(F)═CH—CH═)), 7.34-7.37 (d AB, J=8.6 Hz, 2H,—C₂ H ₂C₂H₂C(NO₂)), 7.62-7.65 (dd, J=7.2, 2.1 Hz, 1H, C(═O)—C═CH),8.04-8.06 (d AB, J=8.6 Hz, 2H, —C₂ H ₂C(NO₂))

¹³C NMR (CDCl₃, 75 MHz), δ 8.87, 8.92, 22.64, 42.90, 67.55, 67.64,94.64, 110.93, 111.25, 121.03, 123.61, 125.35, 125.46, 128.25, 129.03,130.20, 135.38, 137.11, 141.00, 144.64, 162.64, 167.32

IR: 701, 777, 802, 830, 928, 1011, 1065, 1092, 1177, 1227, 1264, 1342,1379, 1447, 1484, 1520, 1605, 1694, 2874, 2928, 3408 cm⁻¹

LCMS (DMSO): Rt=4.05 min (on 5 min column)

HPLC purity (as area %): >98

UV (in EtOH): λ max=267 nm

EI-MS: calculated mass of ion 514.1540 [M+NH₄]⁺, measured mass of ion514.1537 [M+NaH₄]⁺

Rf=0.45 (50% EtOAc/petrol); MP: 64-66° C.

Synthesis of4-chloro-3-(4-chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl)isoindolin-1-one(NU8398) and7-chloro-3-(4-chlorophenyl)-3-hydroxy-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(AW379B, NCL-00016654)

The named compounds were synthesised from a mixture of3-chloro-2-(4-chlorobenzoyl)benzoic acid and6-chloro-2-(4-chlorobenzoyl)benzoic acid (2.15 g, 7.28 mmol) and4-nitrobenzylamine hydrochloride (1.51 g, 8.01 mmol) using GeneralProcedure B, purified by chromatography (Silica; 10%-20% EtOAc/petrol)and obtained as a white solid (NU8398) and a white solid (NCL-00016654)(1.96 g, 63%, ratio of isomers NU8398:NCL-00016654 is 99:1).

Analysis of Major Isomer (NU8398):

¹H NMR (300 MHz, CDCl₃) δ 4.28 (br s, 1H, OH), 4.28 and 4.63 (dd, AB,J=15.4 Hz, 2H, N—CH₂—), 7.19-7.22 (m, 4H, Ar—H), 7.31-7.33 (m, 2H,Ar—H), 7.44-7.49 (m, 2H, Ar—H), 7.72-7.75 (dd, J=3.2, 8.5 Hz, 1H,(C(O)—C═CH—), 7.98-8.00 (m, 2H, —CH—NO₂). ¹³C NMR (CDCl₃, 75 MHz), δ42.37, 90.90, 122.20, 123.22, 123.35, 128.21, 128.68, 129.44, 129.75,131.69, 131.89, 134.14, 135.15, 147.37, 163.60, 163.86. IR: 696, 729,759, 808, 856, 932, 996, 1070, 1092, 1144, 1174, 1271, 1342, 1397, 1462,1518, 1592, 1682, 2026, 2171, 3220 cm⁻¹. LCMS (DMSO): 8.47 min (on 12min column). UV (in EtOH): λ max=268 nm EI-MS: calculated mass of ion429.0403 [M+H]⁺, measured mass of ion 429.0401 [M+H]⁺. Rf: 0.47 (50%EtOAc/petrol). MP: 202-203° C. CHN: C₂₁H₁₄Cl₂N₂O₄+0.2EtOAc requires C,58.59; H, 3.52; N, 6.27. found C, 58.27; H, 3.21; N, 6.48.

Analysis of Minor Isomer (NCL-00016654)

¹H NMR (300 MHz, CDCl₃) δ 3.90 (s, 1H, OH), 4.30-4.66 (dd, AB, J=15.2Hz, 2H, N—CH₂—), 7.17-7.20 (dd, J=1.2, 7.2 Hz, 1H, CH—C(COH)—),7.21-7.24 (m, 4H, —C₆H₄Cl), 7.29-7.35 (dd AB, J=8.8 Hz, 2H, —CC₂ H₂C₂H₂CNO₂), 7.37-7.48 (m, 2H, ArH), 7.94-8.00 (dd AB, J=8.8 Hz, 2H, —CC₂H ₂C₂H₂CNO₂). ¹³C NMR (CDCl₃, 75 MHz), δ 42.88, 90.25, 121.72, 123.70,126.42, 128.16, 129.23, 130.01, 132.04, 132.10, 135.64, 136.58, 145.18,147.78, 151.26, 165.68. IR: 612, 852, 934, 1093, 1343, 1387, 1515, 1604,1686, 2850, 2932, 3078, 3332 cm⁻¹. LCMS (DMSO): R_(T)=3.47 min (on 5 mincolumn), m/z=427 ES⁻. HPLC purity (as area %): >98. UV (in EtOH): λmax=270 nm EI-MS: calculated mass of ion 429.0403 [M+H]⁺, measured massof ion 429.0403 [M+H]⁺. Rf=0.55 (50% EtOAc/petrol). MP: 172-174° C.

Synthesis of2-(4-acetylbenzyl)-3-(4-chlorophenyl)-3-hydroxy-2,3-dihydroisoindol-1-one(NCL-00016045/AW344)

The named compound was synthesised from 2-(4-chlorobenzoyl)-benzoic acid(687 mg, 2.63 mmol) and 4-acetylbenzylammonium trifluoroacetate (630 mg,2.40 mmol) using General Procedure B, purified by chromatography(Silica; 10%-50% EtOAc/petrol) and obtained as white crystals (552 mg,54%).

¹H NMR (300 MHz, CDCl₃) δ 2.44 (s, 3H, CH₃), 4.10-4.60 (dd AB, J=15.1Hz, 2H, N—CH₂), 5.03 (s, 1H, OH), 7.15-7.21 (m, 4H, ArH), 7.24-7.28 (m,3H, ArH), 7.40-7.52 (m, 2H, ArH), 7.57-7.60 (d AB, J=8.2 Hz, 2H, ArH),7.65-7.69 (m, 1H, —CH—C(C═O)). ¹³C NMR (CDCl₃, 75 MHz), δ 26.39, 42.98,91.38, 123.01, 123.59, 128.23, 128.32, 128.76, 129.14, 129.75, 130.64,133.01, 134.69, 136.17, 137.96, 143.66, 149.26, 168.06, 198.01. IR: 696,725, 770, 804, 849, 905, 956, 1013, 1061, 1089, 1200, 1263, 1352, 1398,1427, 1468, 1603, 1666, 2055, 2846, 2934, 3007, 3140 cm⁻¹. LCMS (DMSO):R_(T)=3.78 min (on 5 min column), m/z=392 ES⁺. HPLC purity (as area%): >98. UV (in EtOH): λ max=254 nm. EI-MS: calculated mass of ion392.1048 [M+H]⁺, measured mass of ion 392.1051 [M+H]⁺. Rf=0.19 (25%EtOAc/petrol). MP: 147-150° C.

Synthesis of2-(4-benzoylbenzyl)-3-(4-chlorophenyl)-3-hydroxy-2,3-dihydroisoindol-1-one(AW357, NCL-00014532)

The named compound was synthesised from 2-(4-chlorobenzoyl)-benzoic acid(791 mg, 3.03 mmol) and 4-acetylbenzylammonium trifluoroacetate (640 mg,1.97 mmol) using General Procedure B, purified by chromatography(Silica; 10%-30% EtOAc/petrol) and obtained as white crystals (414 mg,46%).

¹H NMR (300 MHz, CDCl₃) δ 4.20-4.56 (dd AB, J=15.1 Hz, 2H, N—CH₂), 6.01(s, 1H, OH), 7.14-7.20 (m, 4H, ArH), 7.21-7.28 (m, 3H, ArH), 7.31-7.43(m, 6H, ArH), 7.50-7.53 (m, 1H, —CH—C(C═O)), 7.56-7.62 (m, 3H, ArH). ¹³CNMR (CDCl₃, 75

MHz), δ 43.02, 91.46, 123.09, 123.74, 128.37, 128.55, 128.83, 128.99,129.87, 130.15, 130.26, 130.57, 132.65, 133.16, 134.78, 136.45, 137.86,137.97, 143.06, 149.28, 168.18, 196.74. IR: 698, 735, 763, 810, 864,927, 1015, 1063, 1090, 1198, 1275, 1313, 1350, 1400, 1468, 1597, 1655,2023, 2157, 2931, 3065, 3179 cm⁻¹.

LCMS (DMSO): 3.88 min (on 5 min column), m/z=454 ES⁺

HPLC purity (as area %): >97. UV (in EtOH): λ max=259 nm EI-MS:calculated mass of ion 454.1204 [M+H]⁺, measured mass of ion 454.1206[M+H]⁺. Rf=0.45 (25% EtOAc/petrol). MP: 163-164° C.

Synthesis of3-(4-chlorophenyl)-3-hydroxy-2-(4-iodobenzyl)-2,3-dihydroisoindol-1-one(AW345, NCL-00014527)

The named compound was synthesised from 2-(4-chlorobenzoyl)-benzoic acid(1.76 g, 6.75 mmol) and 4-iodobenzylamine hydrochloride (2 g, 7.42 mmol)using General Procedure B, recrystallised from EtOAc/petrol, purified bychromatography (Silica; 6%-50% EtOAc/petrol) and obtained as a whitesolid (1.72 g, 52%).

¹H NMR (300 MHz, CDCl₃) δ 3.37 (s, 1H, OH), 4.17-4.46 (dd AB, J=15.6 Hz,2H, N—CH₂), 6.98-7.01 (d AB, J=7.8 Hz, 2H, CC₂ H ₂C₂H₂C(I)), 7.23-7.32(m, 5H, ArH), 7.50-7.62 (m, 4H, ArH), 7.75-7.78 (m, 1H, —CH—C(C═O)). ¹³CNMR (CDCl₃, 75 MHz), δ 167.15, 149.54, 139.34, 138.32, 136.90, 133.21,133.03, 130.80, 130.70, 129.73, 128.57, 128.38, 123.18, 122.98, 92.38,90.59, 42.32. IR: 694, 719, 762, 790, 845, 926, 1007, 1063, 1093, 1119,1198, 1288, 1352, 1391, 1412, 1467, 1659, 2912, 3175, 3178 cm⁻¹. LCMS(DMSO): RT=4.89 min (on 5 min column), m/z=476 ES⁺. HPLC purity (as area%): >99. UV (in EtOH): λ max=231 nm EI-MS: calculated mass of ion475.9909 [M+H]⁺, measured mass of ion 475.9905 [M+H]⁺. Rf=0.29 (25%EtOAc/petrol). MP: 184-185° C.

Synthesis of3-(4-chlorophenyl)-3-(1-hydroxymethylcyclopropylmethoxy)-2-(4-iodobenzyl)-2,3-dihydroisoindol-1-one(AW350, NCL-00014529)

The named compound was synthesised from NCL-00014527 (499 mg, 1.01 mmol)and 1,1-bis(hydroxymethyl)cyclopropane (0.19 mL, 2.03 mmol) usingGeneral Procedure C, purified by chromatography (Silica; 10%-40%EtOAc/petrol) and obtained as white crystals (487 mg, 83%).

¹H NMR (300 MHz, CDCl₃) δ 0.10-0.15 (m, 2H, cyclopropane CH₂), 0.35-0.42(m, 2H, cyclopropane CH₂), 1.85 (s, 1H, OH), 2.62-2.80 (dd AB, J=9.4 Hz,2H, iso-O—CH ₂—), 3.32-3.48 (m, 2H, —CH ₂OH) 4.12-4.50 (dd AB, J=14.8Hz, 2H, N—CH₂), 6.85-6.93 (d AB, J=8.3 Hz, 2H, CC₂ H ₂C₂H₂C(I)),7.09-7.13 (m, 1H, —CH—CH═C(C═O)), 7.13-7.18 (m, 4H, ArH), 7.44-7.51 (m,4H, ArH), 7.85-7.89 (m, 1H, —CH—C(C═O)). ¹³C NMR (CDCl₃, 75 MHz), δ8.85, 22.56, 42.94, 67.81, 67.93, 94.68, 95.11, 123.30, 124.00, 128.27,128.93, 130.28, 131.54, 133.20, 135.03, 137.45, 137.48, 137.63, 144.90,145.62, 168.40. IR: 679, 758, 792, 812, 847, 877, 922, 953, 1035, 1065,1091, 1177, 1229, 1277, 1318, 1356, 1386, 1418, 1471, 1611, 1684, 1769,2817, 2880, 2943, 3005, 3063, 3508 cm⁻¹. LCMS (DMSO): R_(T)=4.03 min (on5 min column), m/z=560 ES⁺. HPLC purity (as area %): >98. UV (in EtOH):λ max=229 nm EI-MS: calculated mass of ion 560.0484 [M+H]⁺, measuredmass of ion 560.0472 [M+H]⁺. Rf=0.36 (50% EtOAc/petrol). MP: 164-165° C.

Synthesis of3-(4-chlorophenyl)-2-(4-fluorobenzyl)-3-(1-hydroxymethylcyclopropylmethoxy)-2,3-dihydro-isoindol-1-one(AW351, NCL-00014530)

The named compound was synthesised from3-(4-chloro-phenyl)-2-(4-fluorobenzyl)-3-hydroxy-2,3-dihydroisoindol-1-one(372 mg, 1.01 mmol) and 1,1-bis(hydroxymethyl)cyclopropane (0.19 mL,2.03 mmol) using General Procedure C, purified by chromatography(Silica; 10%-40% EtOAc/petrol) and obtained as yellow crystals (238 mg,52%).

¹H NMR (300 MHz, CDCl₃) δ 0.01-0.04 (m, 2H, cyclopropane CH₂), 0.29-0.34(m, 2H, cyclopropane CH₂), 2.05 (s, 1H, OH), 2.57-2.75 (dd AB, J=9.4 Hz,2H, iso-O—CH ₂—), 3.28-3.46 (dd AB, J=11.3 Hz, 2H, —CH ₂OH) 4.11-4.45(dd AB, J=14.8 Hz, 2H, N—CH₂), 6.71-6.77 (m, 2H, ArH), 7.03-7.08 (m, 3H,—ArH), 7.08-7.13 (m, 4H, ArH), 7.39-7.42 (m, 2H, ArH), 7.78-7.82 (m, 1H,—CH—C(C═O)). ¹³C NMR (CDCl₃, 75 MHz), δ 8.80, 21.08, 42.70, 67.75,95.11, 115.06, 123.31, 123.96, 128.30, 128.87, 130.25, 131.20, 131.30,133.16, 133.70, 134.89, 137.76, 145.75, 164.12, 168.47. IR: 760, 808,844, 918, 951, 1008, 1042, 1063, 1086, 1110, 1158, 1222, 1309, 1346,1393, 1429, 1468, 1508, 1602, 1674, 2848, 2929, 3005, 3080, 3396 cm⁻¹.LCMS (DMSO): R_(T)=3.78 min (on 5 min column), m/z=452 ES⁺. HPLC purity(as area %): >96. UV (in EtOH): λ max=254 nm EI-MS: calculated mass ofion 452.1423 [M+H]⁺, measured mass of ion 452.1419 [M+H]⁺. Rf=0.49 (50%EtOAc/petrol). MP: 130-132° C.

Synthesis of2-(4-acetylbenzyl)-3-(4-chlorophenyl)-3-(1-hydroxymethylcyclopropylmethoxy)-2,3-dihydroisoindol-1-one(AW354, NCL-00014531)

The named compound was synthesised from NCL-00016045 (270 mg, 0.69 mmol)and 1,1-bis(hydroxymethyl)cyclopropane (0.13 mL, 1.38 mmol) usingGeneral Procedure C, purified by chromatography (Silica; 20%-70%EtOAc/petrol) and obtained as a yellow oil (108 mg, 33%).

¹H NMR (300 MHz, CDCl₃) δ −0.05-0.03 (m, 2H, cyclopropane CH₂),0.20-0.31 (m, 2H, cyclopropane CH₂), 1.76 (br s, 1H, OH), 2.42 (s, 3H,CH ₃) 2.58-2.70 (dd AB, J=9.4 Hz, 2H, iso-O—CH ₂—), 3.23-3.37 (dd AB,J=11.3 Hz, 2H, —CH ₂OH) 4.20-4.44 (dd AB, J=15.0 Hz, 2H, N—CH₂),6.95-7.10 (m, 5H, —ArH), 7.11-7.16 (m, 2H, ArH), 7.38-7.41 (m, 2H, ArH),7.59-7.64 (m, 2H, ArH), 7.76-7.80 (m, 1H, —CH—C(C═O))

¹³C NMR (CDCl₃, 75 MHz), δ 8.84, 21.10, 26.63, 43.12, 67.85, 69.40,95.09, 123.37, 124.07, 128.31, 128.48, 128.89, 129.61, 130.35, 132.04,133.26, 135.03, 136.75, 137.61, 142.97, 145.65, 168.47, 197.58

IR: 698, 763, 811, 849, 926, 957, 1012, 1061, 1092, 1177, 1267, 1353,1383, 1417, 1466, 1607, 1680, 2875, 2922, 3001, 3368, 3402 cm⁻¹. LCMS(DMSO): 4.14 min (on 5 min column). HPLC purity (as area %): >97. UV (inEtOH): λ max=251 nm. EI-MS: calculated mass of ion 476.1623 [M+H]⁺,measured mass of ion 476.618 [M+H]⁺. Rf=0.21 (50% EtOAc/petrol).

Synthesis of2-(4-benzoylbenzyl)-3-(4-chlorophenyl)-3-(1-hydroxymethylcyclopropylmethoxy)-2,3-dihydroisoindol-1-one(AW360, NCL-00014533)

The named compound was synthesised from NCL-00014532 (142 mg, 0.31 mmol)and 1,1-bis(hydroxymethyl)cyclopropane (0.06 mL, 0.63 mmol) usingGeneral Procedure C, purified by chromatography (Silica; 10%-50%EtOAc/petrol) and obtained as yellow crystals (124 mg, 74%).

¹H NMR (300 MHz, CDCl₃) δ −0.08-0.03 (m, 2H, cyclopropane CH₂),0.25-0.30 (m, 2H, cyclopropane CH₂), 1.94 (br s, 1H, OH), 2.60-2.69 (ddAB, J=9.4 Hz, 2H, iso-O—CH ₂—), 3.23-3.37 (dd AB, J=11.3 Hz, 2H, —CH₂OH) 4.22-4.42 (dd AB, J=15.0 Hz, 2H, N—CH₂), 6.96-7.05 (m, 5H, —C₆ H₄Cl & —CHCHC(C═O—N—), 7.09-7.12 (d AB, J=8.1 Hz, 2H, —N—CH₂—CC₂ H₂C₂H₂C(C(═O)Ph), 7.25-7.42 (m, 7H, ArH), 7.53-7.56 (d AB, J=8.1 Hz, 2H,—N—CH₂—CC₂ H ₂C₂H₂C(C(═O)Ph), 7.73-7.76 (m, 1H, —CH—C(C═O)). ¹³C NMR(CDCl₃, 75 MHz), δ 8.86, 22.65, 43.11, 67.74, 69.30, 95.07, 123.41,124.08, 128.40, 128.56, 128.87, 129.07, 129.26, 130.18, 130.36, 132.04,132.55, 133.28, 134.97, 137.09, 137.72, 138.15, 142.32, 145.69, 168.52,196.28. IR: 699, 727, 764, 813, 858, 922, 1013, 1063, 1092, 1177, 1276,1314, 1383, 1466, 1605, 1656, 1690, 2877, 2921, 3063, 3411 cm⁻¹. LCMS(DMSO): 4.88 min (on 5 min column). HPLC purity (as area %): >94. UV (inEtOH): λ max=261 nm. EI-MS: calculated mass of ion 560.1599 [M+Na]⁺,measured mass of ion 560.1601[M+Na]⁺.

Rf=0.40 (50% EtOAc/petrol). MP: 69-71° C.

Synthesis of3-(4-chlorophenyl)-3-hydroxy-2-naphthalen-1-ylmethyl-2,3-dihydroisoindol-1-one(AW364, NCL-00016046)

The named compound was synthesised from 2-(4-chlorobenzoyl)-benzoic acid(2.24 g, 8.59 mmol) and 1-naphthylmethylamine (1.36 mL, 9.44 mmol) usingGeneral Procedure B, purified by chromatography (Silica; 6%-50%EtOAc/petrol) and obtained as yellow crystals (0.35 g, 10%).

¹H NMR (300 MHz, MeOD) δ 4.90 (s, 1H, OH), 4.90-5.27 (dd AB, J=15.2 Hz,2H, N—CH₂), 6.75-6.79 (m, 2H, ArH), 6.95-6.98 (d AB, J=8.5 Hz, 2H, ArH),7.13-7.27 (m, 3H, ArH), 7.35-7.61 (m, 5H, ArH), 7.72-7.75 (m, 1H,—CH—C(C═O)), 7.86-7.89 (m, 1H, ArH), 8.17-8.19 (m, 1H,N—CH₂—C—C(C)═CH)). ¹³C NMR (MeOD, 75 MHz), δ 41.89, 92.80, 124.27,124.59, 125.20, 126.37, 126.89, 127.38, 129.00, 129.05, 129.32, 129.40,129.96, 131.05, 132.00, 133.36, 134.02, 134.55, 135.03, 135.47, 139.68,151.56, 170.26. IR: 694, 762, 833, 929, 973, 1011, 1060, 1091, 1112,1195, 1268, 1360, 1396, 1468, 1599, 1674, 2070, 2873, 2967, 3053, 3270,3331 cm⁻¹. LCMS (DMSO): R_(T)=4.19 min (on 5 min column), m/z=398 ES⁻.HPLC purity (as area %): >92. UV (in EtOH): λ max=223 nm. EI-MS:calculated mass of ion 400.1099 [M+H]⁺, measured mass of ion 400.1099[M+H]⁺. Rf=0.15 (25% EtOAc/petrol). MP: 75-76° C.

Synthesis of2-β-bromobenzyl)-3-(4-chlorophenyl)-3-hydroxy-2,3-dihydroisoindol-1-one(AW365, NCL-00016047)

The named compound was synthesised from 2-(4-chlorobenzoyl)-benzoic acid(2.24 g, 8.59 mmol) and 3-bromobenzylamine hydrochloride (2.10 g, 9.44mmol) using General Procedure B, recrystallised from EtOAc/petrol andobtained as orange crystals (2.12 g, 58%).

¹H NMR (300 MHz, CDCl₃) δ 4.12-4.33 (dd AB, J=15.0 Hz, 2H, N—CH₂), 4.54(br s, 1H, OH), 6.93-7.04 (m, 2H, —CH—CH═CH—CBr), 7.08-7.10 (m, 1H,ArH), 7.12-7.19 (m, 4H, —C₆ H ₄Cl), 7.20-7.29 (m, 2H, ArH), 7.41-7.53(m, 2H, ArH), 7.64-7.69 (m, 1H, ═CH—C—C(═O)). ¹³C NMR (CDCl₃, 75 MHz), δ42.70, 91.40, 122.52, 122.56, 123.06, 123.95, 127.78, 128.27, 128.89,130.01, 130.15, 130.54, 132.20, 133.36, 135.09, 137.20, 140.35, 149.08,168.05. IR: 664, 696, 721, 764, 802, 837, 879, 926, 976, 1011, 1059,1088, 1191, 1306, 1348, 1399, 1427, 1468, 1572, 1600, 1668, 2875, 2932,3016, 3246 cm⁻¹. LCMS (DMSO): R_(T)=4.32 min (on 5 min column), m/z=428ES⁻. HPLC purity (as area %): >98. UV (in EtOH): λ max=254 nm. EI-MS:calculated mass of ion 428.0047 [M+H]⁺, measured mass of ion 428.0041[M+H]⁺.

Rf=0.42 (50% EtOAc/petrol). MP: 167-170° C.

Synthesis of3-(4-chlorophenyl)-3-(1-hydroxymethylcyclopropylmethoxy)-2-naphthalen-1-ylmethyl-2,3-dihydroisoindol-1-one(AW366, NCL-00016106)

The named compound was synthesised from NCL-00016046 (270 mg, 0.69 mmol)and 1,1-bis(hydroxymethyl)cyclopropane (0.13 mL, 1.38 mmol) usingGeneral Procedure C, purified by chromatography (Biotage SP4; 6%-25%EtOAc/petrol) and obtained as cream crystals (153 mg, 47%).

¹H NMR (300 MHz, CDCl₃) δ −0.35-0.15 (m, 2H, cyclopropane CH), 0.12-0.25(m, 2H, cyclopropane CH), 1.80 (br s, 1H, OH), 2.38-2.70 (dd AB, J=9.4Hz, 2H, CH ₂O—C), 3.18-3.31 (dd AB, J=11.1 Hz, 2H, CH ₂OH), 4.63-5.27(dd AB, J=14.9 Hz, 2H, N—CH₂), 7.02-7.07 (m, 5H, C₆ H ₄Cl & ArH)),7.15-7.30 (m, 2H, ArH), 7.43-7.48 (m, 4H, ArH), 7.68-7.71 (m, 1H,NCH₂C—CHCHCH—C), 7.76-7.79 (m, 1H, —CH—C(C═O)), 7.93-7.97 (m, 1H,N—CH₂—CC(C)═CHCHCHCHC—C), 8.29-8.32 (m, 1H, N—CH₂—C—C(C)═CH)).

¹³C NMR (CDCl₃, 75 MHz), δ 8.58, 8.61, 22.36, 41.25, 67.68, 67.78,95.41, 123.17, 124.09, 124.54, 125.35, 126.02, 126.70, 128.05, 128.50,128.68, 128.85, 129.09, 130.15, 131.92, 132.40, 132.91, 133.19, 134.03,134.49, 137.55, 146.16, 168.32. IR: 696, 767, 810, 839, 923, 945, 1012,1065, 1110, 1271, 1356, 1389, 1467, 1487, 1598, 1685, 2874, 2923, 3004,3051, 3395 cm⁻¹. LCMS (DMSO): RT=3.99 min (on 5 min column), m/z=484ES⁺. HPLC purity (as area %): >95. UV (in EtOH): λ max=223 nm. EI-MS:calculated mass of ion 484.1674 [M+H]⁺, measured mass of ion 484.1673[M+H]⁺. Rf=0.10 (25% EtOAc/petrol). MP: 82-84° C.

Synthesis of2-(3-bromobenzyl)-3-(4-chlorophenyl)-3-(1-hydroxymethylcyclopropylmethoxy)-2,3-dihydro-isoindol-1-one(AW367, NCL-00016107)

The named compound was synthesised from NCL-00016047 (292 mg, 0.68 mmol)and 1,1-bis(hydroxymethyl)cyclopropane (0.13 mL, 1.35 mmol) usingGeneral Procedure C, purified by chromatography (Biotage SP4; 6%-50%EtOAc/petrol) and obtained as a pale yellow oil (253 mg, 73%).

¹H NMR (300 MHz, CDCl₃) δ −0.05-0.05 (m, 2H, cyclopropane CH), 0.22-0.31(m, 2H, cyclopropane CH), 2.33 (br s, 1H, OH), 2.54-2.73 (dd AB, J=9.3Hz, 2H, CH ₂O—C), 3.26-3.32 (dd AB, J=11.0 Hz, 2H, CH ₂OH), 4.10-4.31(dd AB, J=14.9 Hz, 2H, N—CH₂), 6.81-6.87 (m, 1H, —CH—CH═CH—CBr),6.95-7.05 (m, 7H, ArH), 7.09-7.11 (m, 1H, —CH═CH—C—C(═O)), 7.33-7.35 (m,2H, —CH═CH—C—C(O—CH₂—)), 7.71-7.74 (m, 1H, ═CH—C—C(═O)). ¹³C NMR (CDCl₃,75 MHz), δ 8.76, 8.82, 22.58, 42.81, 67.52, 67.60, 94.99, 122.50,123.40, 124.03, 128.19, 128.32, 128.86, 130.02, 130.30, 130.60, 132.00,132.55, 133.25, 134.98, 137.61, 140.02, 145.69, 168.40. IR: 666, 695,711, 761, 812, 838, 927, 1009, 1063, 1090, 1307, 1346, 1380, 1427, 1467,1571, 1595, 1686, 2874, 2920, 3001, 3065, 3429 cm⁻¹. LCMS (DMSO):R_(T)=3.93 min (on 5 min column), m/z=511 ES⁻. HPLC purity (as area%): >98. UV (in EtOH): λ max=203 nm EI-MS: calculated mass of ion512.0623 [M+H]⁺, measured mass of ion 512.0620 [M+H]⁺. Rf=0.36 (50%EtOAc/petrol).

Synthesis of3-hydroxy-2-(4-nitrobenzyl)-3-phenyl-2,3-dihydroisoindol-1-one(AW403/NCL-00016655)

The named compound was synthesised from 2-benzoyl-benzoic acid (0.5 g,2.21 mmol) and 4-nitrobenzylamine hydrochloride (0.46 g, 2.43 mmol)using General Procedure B, recrystallised from EtOAc/petrol and obtainedas a yellow solid (0.56 g, 70%).

¹H NMR (300 MHz, CDCl₃) δ 3.36 (s, 1H, OH), 4.26-4.71 (dd AB, J=15.3 Hz,2H, N—CH₂), 7.22-7.34 (m, 8H, Ar—H), 7.48-7.57 (m, 2H, Ar—H), 7.81-7.84(m, 1H, CH═C—C(═O)), 7.96-8.01 (dd AB, J=8.8 Hz, 2H, C₂H₂C—NO₂). ¹³C NMR(CDCl₃, 75 MHz), δ 41.02, 91.09, 123.65, 124.05, 125.56, 126.65, 127.64,128.93, 129.05, 129.82, 133.48, 138.08, 138.25, 142.31, 145.80, 148.57,168.29. IR: 688, 756, 851, 934, 1055, 1105, 1192, 1283, 1337, 1398,1468, 1514, 1605, 3083, 3181 cm⁻¹. LCMS (DMSO): R_(T)=3.13 min (on 5 mincolumn), m/z=359 ES⁻. HPLC purity (as area %): >99. UV (in EtOH): λmax=269 nm EI-MS: calculated mass of ion 361.1183 [M+H]⁺, measured massof ion 361.1186 [M+H]⁺. Rf=0.35 (50% EtOAc/petrol). MP: 190-191° C.

Synthesis of3-(1-hydroxymethylcyclopropylmethoxy)-2-(4-nitrobenzyl)-3-phenyl-2,3-dihydroisoindol-1-one(AW405/NCL-00016656)

The named compound was synthesised from NCL-00016655 (300 mg, 0.83 mmol)and 1,1-bis(hydroxymethyl)cyclopropane (0.16 mL, 1.67 mmol) usingGeneral Procedure C, purified by chromatography (Biotage SP4; 10%-50%EtOAc/petrol) and obtained as pale yellow crystals (298 mg, 81%).

¹H NMR (300 MHz, CDCl₃) δ −0.02-0.01 (m, 2H, cyclopropane CH₂),0.15-0.25 (m, 2H, cyclopropane CH₂), 2.05 (s, 1H, OH), 2.49-2.65 (m, 2H,C—O—CH₂), 3.20-3.40 (m, 2H, CH ₂OH), 4.20-4.40 (m, 2H, N—CH₂), 6.80-7.10(m, 8H, Ar—H), 7.25-7.35 (m, 2H, Ar—H), 7.70-7.90 (m, 3H, CH═C—C(═O) &C₂H₂C—NO₂)

¹³C NMR (CDCl₃, 75 MHz), δ 8.59, 21.07, 42.08, 66.02, 66.13, 94.53,123.34, 123.83, 126.54, 128.29, 128.81, 129.67, 130.13, 131.52, 133.04,137.95, 138.03, 144.61, 145.46, 146.88, 168.36. IR: 696, 753, 796, 854,910, 938, 1022, 1057, 1103, 1179, 1243, 1278, 1341, 1384, 1466, 1518,1605, 1686, 2853, 2921, 3077, 3414 cm⁻¹.

LCMS (DMSO): RT=3.22 min (on 5 min column). HPLC purity (as area %):>97.

UV (in EtOH): λ max=267 nm EI-MS: calculated mass of ion 445.1758[M+H]⁺, measured mass of ion 445.1757 [M+H]⁺. Rf=0.27 (50%EtOAc/petrol). MP: 58-60° C.

Synthesis of succinic acidmono-{1-[7-chloro-1-(4-chloro-phenyl)-2-(4-nitrobenzyl)-3-oxo-2,3-dihydro-1H-isoindol-1-yloxymethyl]cyclopropylmethyl}ester(AW393/NCL-00016149)

The named compound was synthesised from NU8406 (100 mg, 0.19 mmol),pyridine (0.03 mL, 0.39 mmol), 4-dimethylamino pyridine (5 mg, 0.04mmol) and succinic anhydride (39 mg, 0.39 mmol) in anhydrous THF (10 mL)using General Procedure F, purified by chromatography (Biotage SP4; 50%EtOAc/petrol—20% MeOH/EtOAc) and obtained as white crystals (60 mg,50%).

¹H NMR (300 MHz, CDCl₃) δ 0.20-0.42 (m, 2H, cyclopropane CH₂), 0.45-0.52(m, 2H, cyclopropane CH₂), 2.45-2.52 (br m, 4H, —CH ₂CH ₂CO₂H),2.62-2.93 (dd, AB, J=9.3 Hz, 2H, iso-C—O—CH₂—), 4.00-4.15 (m, 2H, CH₂OCOCH₂CH₂CO₂H), 4.30-4.60 (dd, AB, J=15.2 Hz, 2H, N—CH₂—), 7.01-7.31(m, 6H, Ar—H), 7.42-7.57 (m, 2H, Ar—H), 7.80-8.10 (br m, 4H, ArH andCO₂H). ¹³C NMR (CDCl₃, 75 MHz), δ 9.38, 20.00, 29.20, 29.45, 42.50,67.27, 68.58, 94.68, 122.67, 123.53, 128.51, 128.70, 128.86, 130.06,130.56, 132.31, 134.37, 135.36, 141.10, 142.22, 144.50, 147.67, 167.32,172.27, 176.16. IR: 594, 730, 819, 1076, 1165, 1344, 1521, 1707, 1708,2882, 2929, 3079 cm⁻¹. LCMS (DMSO): RT=3.67 min (on 5 min column),m/z=612 ES⁻

HPLC purity (as area %): >95. EI-MS: calculated mass of ion 613.1139[M+H]⁺, measured mass of ion 613.1139 [M+H]⁺. Rf=0.06 (50%EtOAc/petrol). MP: 42-44° C.

Synthesis of succinic acidmono-{1-[7-chloro-1-(4-chlorophenyl)-2-(4-cyanobenzyl)-3-oxo-2,3-dihydro-1H-isoindol-1-yloxymethyl]cyclopropylmethyl}ester(AW417/NCL-00016659)

The named compound was synthesised from NCL-00010492 (110 mg, 0.22mmol), pyridine (36 mL, 0.45 mmol), 4-dimethylamino pyridine (5 mg, 0.04mmol) and succinic anhydride (45 mg, 0.45 mmol) in anhydrous THF (10 mL)using General Procedure F, purified by chromatography (Biotage SP4; 50%EtOAc/petrol—20% MeOH/EtOAc) and obtained as white crystals (20 mg,15%).

¹H NMR (300 MHz, CDCl₃) δ −0.27-0.20 (m, 2H, cyclopropane CH₂),0.29-0.36 (m, 2H, cyclopropane CH₂), 2.30-2.42 (br m, 4H, —CH ₂CH₂CO₂H), 2.55-2.66 (dd, AB, J=9.3 Hz, 2H, iso-C—O—CH₂—), 3.85-3.90 (m,2H, CH ₂OCOCH₂CH₂CO₂H), 4.05-4.32 (dd, AB, J=15.2 Hz, 2H, N—CH₂—),6.80-6.92 (br m, 5H, —C₆ H ₄Cl and CO₂H), 6.93 (d AB, J=8.3 Hz, 2H, CC₂H ₂C₂H₂CCN), 7.18 (d AB, J=8.3 Hz, 2H, CC₂ H ₂C₂H₂CCN), 7.23-7.25 (dd,J=8.0, 0.9 Hz, 1H, —CH—CH═C(Cl)—C—C(OCH₂—), 7.28-7.32 (m, 1H,—CH═C(Cl)—C—C(OCH₂—)), 7.62-7.65 (dd, J=7.4, 0.9 Hz, 1H, C(═O)—C═CH).¹³C NMR (CDCl₃, 75 MHz), δ 9.01, 9.07, 19.49, 28.90, 28.99, 29.70,42.28, 66.69, 68.23, 94.20, 111.07, 118.54, 122.37, 128.34, 129.58,130.03, 131.88, 132.03, 133.86, 134.03, 134.71, 134.84, 140.60, 142.29,167.08, 172.22, 176.80. IR: 728, 761, 814, 853, 928, 1009, 1074, 1161,1207, 1373, 1458, 1719, 1730, 2227, 2857, 2926, 3005, 3071 cm⁻¹.

LCMS (DMSO): RT=3.41 min (on 5 min column), m/z=592 ES⁻. HPLC purity (asarea %): >96. UV (in EtOH): λ max=226.5 nm. EI-MS: calculated mass ofion 593.1241 [M+NH₄]⁺, measured mass of ion 593.1240 [M+NH₄]⁺. Rf=0.05(50% EtOAc/petrol). MP: 72-74° C.

Synthesis of succinic acidmono-{1-[2-(4-bromobenzyl)-7-chloro-1-(4-chlorophenyl)-3-oxo-2,3-dihydro-1H-isoindol-1-yloxymethyl]cyclopropylmethyl}ester(AW436, NCL-00016653)

The named compound was synthesised from NCL-00010493 (150 mg, 0.27mmol), pyridine (44 μL, 0.55 mmol), 4-dimethylamino pyridine (7 mg, 0.05mmol) and succinic anhydride (55 mg, 0.55 mmol) in anhydrous THF (10 mL)using General Procedure I, purified by chromatography (Biotage SP4; 50%EtOAc/petrol —EtOAc) and obtained as a brown oil (93 mg, 53%).

¹H NMR (300 MHz, CDCl₃) δ −0.04-0.18 (m, 2H, cyclopropane CH₂),0.27-0.35 (m, 2H, cyclopropane CH₂), 2.37-2.46 (br m, 4H, —CH ₂CH₂CO₂H), 2.52-2.69 (dd, AB, J=9.3 Hz, 2H, iso-C—O—CH₂—), 3.74-4.03 (ddAB, J=11.4 Hz, 2H, CH ₂OCOCH₂CH₂CO₂H), 4.05-4.17 (dd, AB, J=14.9 Hz, 2H,N—CH₂—), 6.78 (d AB, J=8.4 Hz, 2H, —C₂ H ₂C₂H₂Br), 6.90-7.01, (m, 4H,CC₂H₄Cl), 7.06 (d AB, J=8.4 Hz, 2H C₂ H ₂—C(Br)), 7.25-7.27 (dd, J=7.9,0.8 Hz, 1H, —CH—CH═C(Cl)—C—C(OCH₂—), 7.30-7.33 (m, 1H,—CH═C(Cl)—C—C(OCH₂—) 7.65-7.67 (dd, J=7.4, 0.8 Hz, 1H, C(═O)—C═CH) 9.00(br s, 1H, COOH). ¹³C NMR (CDCl₃, 75 MHz), δ 8.92, 8.98, 19.34, 28.87,28.90, 42.21, 66.60, 68.37, 94.45, 121.33, 122.28, 128.33, 128.43,129.91, 130.78, 131.21, 131.88, 133.86, 134.02, 134.67, 134.84, 135.97,140.75, 166.97, 172.14, 177.17. IR: 760, 817, 926, 1009, 1070, 1159,1350, 1387, 1462, 1487, 1588, 1704, 1730, 2854, 2921 cm⁻¹. LCMS (DMSO):RT=3.64 min (on 5 min column), m/z=648 ES⁺. HPLC purity (as area%): >96. UV (in EtOH): λ max=222.5 nm EI-MS: calculated mass of ion663.0659 [M+NH₄]⁺, measured mass of ion 663.0653 [M+NH₄]⁺

Rf=0.08 (50% EtOAc/petrol).

3-(4-chlorophenyl)-3-hydroxy-2-(4-methylbenzyl)isoindolin-1-one (tjb14/02)

The named compound was synthesised from 2-(4-chlorobenzoyl)-benzoic acid(2.0 g, 7.6 mmol) and 4-methyl benzylamine (1.07 mL, 8.4 mmol) usingGeneral Procedure B, purified by chromatography (Biotage SP4; 10-80%EtOAc/hexane) and obtained as a white solid (2.045 g, 72%).

¹H NMR (500 MHz, CDCl₃) δ 7.74-7.77 (1H, m, 7-H), 7.53-7.59 (2H, m, 5 &6-H), 7.28-7.31, (3H, m, Ar—H), 7.24,-7.26 (2H, m, Ar—H), 7.05 (2H, d,J=8.0, Ar—H), 6.96 (2H, d, J=8.0, Ar—H), 4.41 (1H, d, J=15.4, 2-CH),4.20, (1H, d, J=15.4, 2-CH′), 2.22 (3H, s, CH₃). ¹³C NMR (125 MHz,CDCl₃) δ 166.7, 149.1, 139.0, 135.5, 135.0, 132.7, 132.6, 130.3, 129.4,128.3, 128.2, 128.0, 127.9, 122.8, 122.6, 90.2, 42.1, 20.6. Found;364.1101 [M+H]; C₂₂H₁₉NO₂Cl, requires 364.1099.

3-(4-chlorophenyl)-3-hydroxy-2-(4-methoxybenzyl)isoindolin-1-one (tjb16/02)

The named compound was synthesised from 2-(4-chlorobenzoyl)-benzoic acid(2.0 g, 7.6 mmol) and 4-methoxy benzylamine (0.542 mL, 4.18 mmol) usingGeneral Procedure B, purified by chromatography (Biotage SP4; 10-80%EtOAc/hexane) and obtained as a white solid (0.830 g, 57%).

¹H NMR (500 MHz, CDCl₃) δ 7.78-7.80 (1H, m, 7-H), 7.57-7.61 (2H, m, 5 &6-H), 7.29-7.33 (3H, m, Ar—H), 7.25-7.27 (2H, m, Ar—H), 7.10-7.13 (2H,m, Ar—H), 6.73-6.76 (2H, m, Ar—H), 4.40 (1H, d, J=15.2, 2-CH), 4.26 (1H,d, J=15.2, 2-CH′), 3.72 (3H, s, CH₃). ¹³C NMR (125 MHz, CDCl₃) δ 166.7,157.9, 149.1, 139.0, 132.6, 132.5, 130.3, 130.0, 129.4, 129.3, 128.2,128.0, 122.8, 122.5, 113.190.1, 55.0, 41.72. Found; 380.1055 [M+H];C₂₂H₁₉NO₃Cl, requires 380.1048.

3-(4-chlorophenyl)-3-(1′-hydroxy-2′-cyclopropyl-3′-methoxy)-2-(4-methylbenzyl)isoindolin-1-one(NCL-00016865)

The named compound was synthesised from3-(4-chlorophenyl)-3-hydroxy-2-(4-methylbenzyl)isoindolin-1-one (300 mg,0.83 mmol) and 1,1-bis(hydroxymethyl)cyclopropane (0.16 mL, 1.67 mmol)using General Procedure C, purified by chromatography (Biotage SP4;10%-80% EtOAc/n-hexane) and obtained as a glassy solid (187 mg, 54%).

¹H NMR (500 MHz, CDCl₃) δ 7.82-7.84 (1H, m, 7-H), 7.41-7.45 (2H, m, 5 &6-H), 7.15 (4H, s, Ar—H), 7.04-7.07 (3H, m, 4 & Ar—H), 6.93 (2H, d,J=7.8, Ar—H), 4.60 (1H, d, J=14.8, 2-CH), 3.95 (1H, d, J=14.8, 2-CH′),3.31 (1H, d, J=11.3, 1′-H), 3.25 (1H, d, J=11.3, 1′-H′), 2.64 (1H, d,J=9.5, 3′-H), 2.55 (1H, d, J=9.5, 3′-H′), 2.21 (3H, s, CH₃), 1.54 (1H,br s, 1′-OH), 0.27-0.30 (2H, m, H₂), −0.02-0.03 (2, m, H₂′). ¹³C NMR(125 MHz, CDCl₃) δ 168.2, 145.3, 137.3, 137.1, 134.5, 134.4, 132.8,131.7, 129.9, 129.2, 128.9, 128.7, 127.9, 123.7, 122.8, 95.0, 68.0,67.7, 42.9, 29.7, 22.1, 21.0, 8.6. Found; 448.1673 [M+H]; C₂₇H₂₇NO₃Cl,requires 448.1674.

3-(4-chlorophenyl)-3-(1′-hydroxy-1-cyclopropyl-3′-methoxy)-2-(4-methoxybenzyl)isoindolin-1-one(NCL-00016866)

The named compound was synthesised from3-(4-chlorophenyl)-3-hydroxy-2-(4-methoxybenzyl)isoindolin-1-one (300mg, 0.79 mmol) and 1,1-bis(hydroxymethyl)cyclopropane (0.12 mL, 1.58mmol) using General Procedure C, purified by chromatography (BiotageSP4; 10%-80% EtOAc/n-hexane) and obtained as a glassy solid (181 mg,53%).

¹H NMR (500 MHz, CDCl₃) δ 7.75-7.86 (1H, m, 7-H), 7.35-7.39 (2H, m, 5 &6-H), 7.08 (4H, m, Ar—H), 6.99-7.02 (3H, m, 4 & Ar—H), 6.57-6.60 (2H, m,Ar—H), 4.45 (1H, d, J=14.8, 2-CH), 3.97 (1H, d, J=14.8, 2-CH′), 3.62(3H, s, CH₃), 3.32 (1H, d, J=11.3, 1′-H), 3.22 (1H, d, J=11.3, 1′-H′),2.62 (1H, d, J=9.4, 3′-H), 2.53 (1H, d, J=9.4, 3′-H′), 1.53 (1H, br s,1′-OH), 0.23-0.28 (2H, m, H₂), −0.02-0.02 (2H, m, H₂′).

¹³C NMR (125 MHz, CDCl₃) δ 168.1, 158.9, 145.3, 137.2, 134.4, 132.8,131.7, 130.5, 129.9, 129.8, 128.6, 127.9, 123.6, 122.7, 113.6, 94.9,68.0, 67.7, 55.3, 42.5, 22.1, 8.7, 8.6. Found; 464.1619 [M+H];C₂₇H₂₇NO₃Cl, requires 464.1623.

3-(4-chlorophenyl)-3-(1′-hydroxy-2′-cyclopropyl-3′-methoxy)-2-(4-carboxamidebenzyl)isoindolin-1-one(NCL-00016867)

To a solution of NCL-00010492 (200 mg, 0.436 mmol) in t-BuOH (8.48 mL)at 50° C. was added finely powdered KOH (647 mg). The resultingsuspension was stirred at this temperature until TLC (10% MeOH/DCM)indicated the complete consumption of the starting material (3 h). Thehot reaction mixture was filtered through Celite® and the pad rinsedwith several portions of THF. The filtrate was partitioned between EtOAc(10 mL) and H₂O (10 mL) and the organic layer separated; the aqueouslayer was extracted with EtOAc (2×10 mL). The combined organic layerswere washed with brine (15 mL), dried (Na₂SO₄), filtered andconcentrated in vacuo. The residue was dissolved in THF before theaddition of a minimum amount of silica and the resulting suspension wasconcentrated in vacuo. Purification by flash column chromatography onsilica gel, eluting with 2-10% MeOH in DCM afforded the title compoundas a glassy solid (62 mg, 30%).

¹H NMR (500 MHz, CDCl₃) δ 7.77-7.80 (1H, m, 7-H), 7.51 (2H, d, J=8.2,Ar—H), 7.38-7.42 (2H, m, 5 & 6-H), 7.13-7.15 (2H, m, Ar—H), 7.06 (4H, m,Ar—H), 7.00-7.02 (1H, m, 4-H), 6.11, (1H, br s, N—H), 5.71 (1H, br s,N—H′), 4.48 (1H, d, J=15.0, 2-CH), 4.16 (1H, d, J=15.0, 2-CH′), 3.31(1H, d, J=10, 1′-H), 3.25 (1H, d, J=10, 1′-H′), 2.62 (2H, s, 3′-H₂),1.66 (1H, br s, 1′-OH) 0.24-0.34 (2H, m, H₂), −0.01-0.04 (2H, m, H₂).¹³C NMR (125 MHz, CDCl₃) δ 169.2, 168.3, 145.1, 141.5, 137.0, 134.6,133.1, 132.3, 131.4, 130.1, 129.3, 128.7, 127.9, 127.3, 123.8, 123.0,94.8, 67.6, 67.4, 50.8, 42.7, 22.2, 8.6. Found; 499.1395 [M+Na];C₂₇H₂₅N₂O₄ClNa, requires 499.1395.

Synthesis of3-(4-chlorophenyl)-2-(4-fluorobenzyl)-3-hydroxy-2,3-dihydroisoindol-1-one(AW349, NCL-0014528)

The named compound was synthesised from 2-(4-chlorobenzoyl)benzoic acid(2.24 g, 8.59 mmol) and 4-fluorobenzylamine (1.08 mL, 9.44 mmol) usingGeneral Procedure B, recrytsllised from EtOAc/petrol, purified bychromatography (Biotage SP4; Silica; 10%-30% EtOAc/petrol) and obtainedas white crystals (1.57 g, 50%).

¹H NMR (300 MHz, CDCl₃) δ 4.40-4.53 (dd AB, J=15.2 Hz, 2H, N—CH₂), 4.58(s, 1H, OH), 6.80-6.89 (m, 2H, CC₂ H ₂C₂H₂C(F)), 7.14-7.29 (m, 7H, ArH),7.50-7.60 (m, 2H, ArH), 7.80-7.86 (m, 1H, —CH—C(C═O)). ¹³C NMR (CDCl₃,75 MHz), δ 43.58, 92.63, 115.85, 116.13, 124.37, 124.52, 129.65, 129.82,131.12, 131.91, 132.02, 134.55, 135.57, 135.70, 139.95, 151.19, 165.35,170.25. IR: 696, 723, 766, 810, 835, 922, 1011, 1059, 1088, 1191, 1223,12658, 1323, 1356, 1397, 1466, 1506, 1602, 1664, 2021, 2851, 2925, 3301cm⁻¹. LCMS (DMSO): RT=3.71 min (on 5 min column), m/z=368 ES⁺. HPLCpurity (as area %): >98. UV (in EtOH): λ max=254 nm EI-MS: calculatedmass of ion 368.0848 [M+H]⁺, measured mass of ion 368.0846 [M+H]⁺.Rf=0.49 (50% EtOAc/petrol). MP: 149-150° C.

Synthesis of3-(4-fluorophenyl)-3-hydroxy-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(AW408/NCL-00016657)

The named compound was synthesised from 2-(4-fluorobenzoyl)-benzoic acid(2.10 g, 8.60 mmol) and 4-nitrobenzylamine hydrochloride (1.78 g, 9.46mmol) using General Procedure B, recrystallised from EtOAc/petrol,purified by chromatography (Biotage; silica, 10%-40% EtOAc/petrol) andobtained as a yellow solid (1.01 g, 31%).

¹H NMR (300 MHz, CDCl₃) δ 4.21 (s, 1H, OH), 4.17-4.15 (dd AB, J=15.4 Hz,2H, N—CH₂), 6.84-6.88 (m, 2H, ArH), 7.19-7.25 (m, 5H, Ar—H), 7.39-7.48(m, 2H, Ar—H), 7.66-7.88 (m, 1H, CH═C—C(═O)), 7.88-7.91 (dd AB, J=8.8Hz, 2H, C₂H₂C—NO₂). ¹³C NMR (CDCl₃, 75 MHz), δ 42.20, 91.07, 115.33,115.51, 122.85, 123.28, 123.52, 128.28, 128.35, 129.42, 129.87, 133.27,133.78, 145.30, 146.90, 148.72, 167.92. IR: 660, 692, 697, 760, 795,814, 849, 892, 932, 1011, 1059, 1095, 1153, 1194, 1219, 1271, 1337,1395, 1421, 1468, 1509, 1601, 1668, 3078, 3282 cm⁻¹. LCMS (DMSO):RT=3.18 min (on 5 min column), m/z=377 ES⁻. HPLC purity (as area%): >95. UV (in EtOH): λ max=266 nm EI-MS: calculated mass of ion379.1089 [M+H]⁺, measured mass of ion 379.1084 [M+H]⁺. Rf=0.46 (50%EtOAc/petrol). MP: 183-185° C.

Synthesis of3-(4-fluorophenyl)-3-(1-hydroxymethylcyclopropylmethoxy)-2-(4-nitrobenzyl)-2,3-dihydro-isoindol-1-one(AW413/NCL-00016896)

The named compound was synthesised from NCL-00016657 (380 mg, 1.01 mmol)and 1,1-bis(hydroxymethyl)cyclopropane (0.19 mL, 2.03 mmol) usingGeneral Procedure C, purified by chromatography (Biotage SP4; silica;25%-50% EtOAc/petrol) and obtained as a pale yellow oil (327 mg, 70%).R_(f)=0.29 (50:50 EtOAc:petrol). mp 57-58° C. λ_(max) (CH₃OH)/nm=266.IR: 711, 727, 762, 802, 818, 853, 918, 1014, 1061, 1098, 1157, 1183,1229, 1278, 1341, 1383, 1411, 1468, 1514, 1602, 1690, 2876, 2924, 3079,3393 cm⁻¹. ¹H NMR: (300 MHz, CDCl₃) δ 0.14-0.31 (m, 2H, cyclopropaneCH₂), 0.43-0.51 (m, 2H, cyclopropane CH₂), 2.17 (br s, 1H, OH),2.82-2.91 (dd AB, J=9.4 Hz, 2H, CH₂O—C), 3.49-3.57 (dd AB, J=11.4 Hz,2H, CH₂OH), 4.48-4.62 (dd AB, J=15.2 Hz, 2H, N—CH₂), 6.81-6.92 (m, 2H,ArH), 7.17-7.28 (m, 3H, ArH), 7.31-7.35 (dd AB, J=8.7 Hz, 2H,CC₂H2C₂H₂C(NO₂)), 7.52-7.60 (m, 2H, ArH), 7.90-7.96 (m, 1H, CH═C—C(═O)),7.98-8.02 (dd AB, J=8.7 Hz, 2H, CC₂H₂C₂H₂C(NO₂)). ¹³C NMR: (CDCl₃, 75MHz) δ 8.85, 8.90, 22.66, 42.63, 67.65, 67.66, 94.92, 115.50, 123.54,124.06, 128.74, 128.85, 130.16, 130.43, 131.81, 133.45, 134.67, 145.10,145.71, 147.61, 161.51, 168.59. LCMS (DMSO): R_(T)=3.46 min (on 5 mincolumn), m/z=462 ES⁺. HPLC purity (as area %): >99. HRMS (EI): m/zCalcd. for ion: 463.1664 [M+H]⁺. Found: 463.1662 [M+H]⁺.

Synthesis of 3-chloro-2-(4-chlorobenzoyl)-4-fluorobenzoic acid (AW442)

n-Butyl lithium (2.5M solution, 5.27 mL, 13.18 mmol) was added to astirred solution of diisopropylamine (1.93 mL, 13.75 mmol) in anhydrousTHF (25 mL) at −75° C. under a nitrogen atmosphere, and maintained at−30° C. for a further 1 h to produce lithium diisopropylamide (LDA).After re-cooling to −75° C., a solution of 3-chloro-4-fluorobenzoic acid(1 g, 5.73 mmol) in THF (20 mL) was added over 1 h, and stiffingcontinued overnight at −75° C. under nitrogen. A solution of methyl4-chlorobenzoate (1.95 g, 11.46 mmol) in THF (20 mL) was added over 10min, stiffing was continued at −70° C. for 2 h and then at RT for 4 h.Water (30 mL) was added and the aqueous layer was washed with ether(3×50 mL), acidified with 1M HCl, extracted with DCM (3×50 mL), driedover MgSO₄ and concentrated in vacuo to afford a yellow solid. Partialpurification was attempted with chromatography (Biotage, silica, 50%EtOAc/petrol to 20% MeOH/EtOAc). The crude product (0.60 g, 34%) wasused in the next step without further purification. R_(f)=0.05 (50:50EtOAc:petrol). mp=188-190° C. λ_(max) (CH₃OH)/nm=260. IR: 706, 749, 785,843, 901, 958, 987, 1003, 1090, 1166, 1254, 1395, 1487, 1562, 1586,1671, 1770, 2855, 2924, 3398 cm⁻¹.

¹H NMR: (300 MHz, MeOD) δ 7.27-7.32 (dd AB, J=8.5 Hz, 2H,CC₂H₂C₂H₂C(Cl)), 7.39-7.42 (m, 1H, CHC(F)C(Cl)), 7.60-7.66 (dd AB, J=8.5Hz, 2H, CC₂H₂C₂H₂C(Cl)), 8.17 (d, J=8.30 Hz, 1H, CHCHC(F)C(Cl), 13.60(br s, 1H, CO₂H). ¹³C NMR (MeOD, 75 MHz), δ 119.90, 124.16, 132.022,132.46, 133.89, 134.91, 135.87, 139.02, 163.04, 165.05, 197.46. LCMS(DMSO): RT=3.43 min (on 5 min column), m/z=311 ES⁻.

Synthesis of4-chloro-3-(4-chlorophenyl)-5-fluoro-3-hydroxy-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(AW448/NCL-00016897)

The named compound was synthesised from crude3-chloro-2-(4-chlorobenzoyl)-4-fluorobenzoic acid (150 mg, 0.479 mmol)and 4-nitrobenzylamine hydrochloride (181 mg, 0.958 mmol) using GeneralProcedure B, purified by chromatography (Biotage SP4; silica; 10-40%EtOAc/petrol), recrystallised from EtOAc/petrol and obtained as a whitesolid (0.04 mg, 2%). R_(f)=0.43 (50:50 EtOAc:petrol). mp=289-291° C.λ_(max) (CH₃OH)/nm=265. IR: 672, 705, 738, 816, 853, 897, 959, 1012,1090, 1148, 1204, 1256, 1345, 1401, 1423, 1516, 1585, 1676, 2859, 2974,3210 cm⁻¹.

¹H NMR: (300 MHz, CDCl₃) δ 4.30 (br s, 1H, OH), 4.31-4.70 (dd, J=15.5Hz, 2H, NCH₂), 6.90 (dd AB, J=8.1 Hz, 2H, —CC₂H₂C₂H₂C(Cl)), 7.22-7.26(m, 2H, —CC₂H₂C₂H₂C(Cl)), 7.38 (dd AB, J=8.4 Hz, 2H, —CC₂H₂C₂H₂C(NO₂)),7.64 (d, J=8.0 Hz, 1H, CHC(F)C(Cl)), 7.99 (d, J=8.4 Hz, 1H,CHCHC(F)C(Cl)), 8.05 (dd AB, J=8.4 Hz, 2H, —CC₂H₂C₂H₂C(NO₂)). ¹³C NMR:(CDCl₃, 75 MHz) δ 42.73, 90.17, 123.42, 123.54, 126.35, 128.08, 128.46,128.93, 129.38, 130.55, 131.80, 131.80, 134.71, 141.26, 144.16, 144.57,167.99, 168.46. LCMS (DMSO): RT=4.06 (on 5 min column), m/z=445 ES⁻.HPLC purity (as area %): >95. HRMS (EI): m/z Calcd. for ion: 445.0164[M−H]. Found: 445.0159 [M−H].

Synthesis of4-chloro-3-(4-chlorophenyl)-3-((1-(hydroxymethyl)cyclopropyl)methoxy)-2-(4-nitrobenzyl)isoindolin-1-one(NU8406A/NCL-00013774 and NU8406B/NCL-00013775)

The named compound was synthesised from NU8398 (433 mg, 1.01 mmol) and1,1-bis(hydroxymethyl)cyclopropane (0.19 mL, 2.03 mmol) using GeneralProcedure C, purified by chromatography (Biotage SP4; silica; 20%-50%EtOAc/petrol) and obtained as yellow crystals (321 mg, 62%). R_(f)=0.30(50:50 EtOAc:petrol). mp 76-77° C. λ_(max) (CH₃OH)/nm=267. IR: 696, 759,816, 853, 930, 1011, 1074, 1144, 1171, 1234, 1341, 1384, 1428, 1462,1489, 1519, 1699, 2872, 2923, 3422 cm⁻¹.

¹H NMR: (300 MHz, CDCl₃) δ 0.21-0.42 (m, 2H, cyclopropane CH₂),0.47-0.54 (m, 2H, cyclopropane CH₂), 2.12 (br s, 1H, OH), 2.89-3.05 (m,2H, C—O—CH₂—), 3.52-3.61 (m, 2H, CH₂OH), 4.30-4.59 (dd, AB, J=15.2 Hz,N—CH₂—), 7.15-7.18 (m, 4H, ArH), 7.28-7.33 (m, 2H, ArH), 7.48-7.58 (m,2H, ArH), 7.87-7.89 (dd, J=7.1, 1.1 Hz, 1H, —C(O)—C═CH—), 7.98-8.01 (m,2H, —CH—NO₂). ¹³C NMR: (CDCl₃, 75 MHz) δ 8.84, 8.90, 22.59, 42.54,67.58, 68.10, 94.71, 122.63, 123.56, 128.65, 128.77, 130.09, 130.40,132.28, 134.37, 135.27, 135.55, 135.57, 141.10, 144.62, 147.64, 167.04.HPLC purity (as area %): >92. HRMS (ED: m/z Calcd. for ion: 530.1244[M+NH₄]⁺. Found: 530.1242 [M+NH₄]⁺. Anal. Calcd. for C₂₆H₂₂Cl₂N₂O₅: C,60.83; H, 4.32; N, 5.46%. Found: C, 60.68; H, 4.30; N, 5.40%.

Separation of enantiomers was achieved by chiral preparative HPLC(Daicel Chiralpak AD-H 250×10 mm; Hexane/Ethanol (4:1))

NU8406A/NCL-00013774 (White Crystals)

Optical rotation: Specific rotation [α]=−4.98° (at 22.4° C.,wavelength=589 nm, tube length=0.25 dm, concentration=0.402 g/100 mL).

NU8406B/NCL-00013775 (White Crystals)

Optical Rotation: Specific rotation [α]=+4.85° (at 22.6° C.,wavelength=589 nm, tube length=0.25 dm, concentration=0.412 g/100 mL).

Synthesis of3-(4-chlorophenyl)-3-(2-hydroxymethylallyloxy)-2-(4-nitrobenzyl)-2,3-dihydroisoindol-1-one(AW468/NCL-00016895)

The named compound was synthesised from NU8260 (400 mg, 1.01 mmol) and2-methylene-1,3-propanediol (0.17 mL, 2.03 mmol) using General ProcedureC, purified by chromatography (Biotage SP4; silica; 10%-40%EtOAc/petrol) and obtained as a yellow oil (342 mg, 73%). R_(f)=0.24(50:50 EtOAc:petrol). λ_(max) (CH₃OH)/nm 268. IR: 702, 764, 808, 853,922, 969, 1011, 1058, 1092, 1177, 1278, 1341, 1381, 1425, 1466, 1489,1519, 1603, 1689, 2859, 2922, 3080, 3407 cm⁻¹.

¹H NMR (300 MHz, CDCl₃) δ 2.46 (br s, 1H, OH), 3.20-3.40 (dd, J=11.9 Hz,2H, iso-OCH₂), 4.00 (s, 2H, CH₂OH), 4.31-4.61 (dd, J=15.1 Hz, 2H, NCH₂),4.88 (s, 1H, C═CH), 5.04 (s, 1H, C═CH), 7.12-7.15 (m, 1H, ArH),7.15-7.21 (dd AB, J=8.7 Hz, 4H, C₆H₄Cl), 7.33-7.36 (dd AB, J=8.7 Hz,2H,)), 7.49-7.53 (m, 2H, ArH), 7.87-7.91 (m, 1H, ArH), 7.95-7.98 (dd AB,2H, CC₂H₂C₂H₂C(NO₂)). ¹³C NMR (CDCl₃, 75 MHz), δ 42.37, 63.63, 63.73,94.80, 112.29, 123.27, 123.34, 123.86, 127.92, 128.69, 130.00, 130.29,131.10, 133.23, 134.78, 136.74, 144.17, 144.57, 144.73, 147.03, 168.32.LCMS (DMSO): Rt=3.74 min (on 5 min column), m/z=465 (ES⁺). HPLC purity(as area %): >98. HRMS (EI): m/z Calcd. for ion: 464.113900 [M]. Found:464.115410 [M].

It is, of course, to be understood that the invention is not intended tobe restricted to the details of the above embodiments which aredescribed by way of example only.

REFERENCES

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1-40. (canceled)
 41. A compound of formula I:

or a pharmaceutically acceptable salt thereof, wherein in formulae I: Xis O; R¹ is selected from substituted or unsubstituted aralkyl,substituted alkyl, propyl, substituted or unsubstituted hydroxyalkyl,substituted or unsubstituted alkylamine, substituted or unsubstitutedaryl, substituted or unsubstituted heteroaryl, and substituted orunsubstituted heteroaralkyl; R² is selected from substituted orunsubstituted alkenyl, substituted or unsubstituted alkynyl, substitutedor unsubstituted branched hydroxyalkyl, substituted or unsubstitutedcycloalkyl having 6 ring carbon atoms or greater, substituted orunsubstituted cycloalkenyl, hydroxyalkylaralkyl,hydroxyalkylheteroaralkyl, and a carboxylic acid-containing group; R³ isselected from substituted or unsubstituted alkyl, substituted orunsubstituted hydroxyalkyl, substituted or unsubstituted alkylamine,substituted or unsubstituted alkoxy, substituted or unsubstituted aryl,substituted or unsubstituted heteroaryl, substituted or unsubstitutedaralkyl, and substituted or unsubstituted heteroaralkyl; and R⁴-R⁷represents groups R⁴, R⁵, R⁶ and R⁷ which are independently selectedfrom hydrogen, halo, hydroxy, substituted or unsubstituted alkyl,substituted or unsubstituted hydroxyalkyl, substituted or unsubstitutedalkenyl, substituted or unsubstituted alkynyl, substituted orunsubstituted heteroaryl, substituted or unsubstituted heteroaralkyl,substituted or unsubstituted alkylamine, substituted or unsubstitutedalkoxy, trifluoromethyl, amino, nitro, carboxyl, carbonyl,methylsulfone, trifluoromethylsulfone, cyano and substituted orunsubstituted sulfonamide.
 42. A compound according to claim 41, whereinR¹ is selected from substituted or unsubstituted aryl, and substitutedor unsubstituted aralkyl; and/or R² is selected from acetyl,hydroxyalkenyl, hydroxyalkynyl, branched 5-carbon hydroxyalkyl,hydroxycycloalkyl, hydroxycycloalkenyl, hydroxymethylcycloalkyl,hydroxymethylcycloalkylmethylene, and hydroxyl alkylbenzyl; and/or R³ isselected from substituted or unsubstituted aryl, and substituted orunsubstituted aralkyl.
 43. A compound according to claim 41, wherein R²is selected from hydroxypropyl, hydroxybutyl, hydroxybutenyl,hydroxycyclopentenyl, hydroxycyclohexenyl and n-propylamine.
 44. Acompound according to claim 41, wherein R¹ is substituted benzyl.
 45. Acompound according to claim 41, wherein R¹ is 4-nitrobenzyl,4-chlorobenzyl, 4-bromobenzyl, cyanobenzyl, or 4-iodobenzyl.
 46. Acompound according to claim 41, wherein R³ is substituted orunsubstituted phenyl.
 47. A compound according to claim 41, wherein R³is 4-chlorophenyl or 4-fluorophenyl.
 48. A compound according to claim41, wherein at least one of R⁴-R¹ is a chlorine atom.
 49. A compoundaccording to claim 41, or a pharmaceutically acceptable salt thereof,wherein: R¹ is selected from substituted aryl, substituted heteroaryl,substituted aralkyl, and substituted heteroaralkyl.
 50. A compoundaccording to claim 41, wherein R¹ is substituted aralkyl, R² is acyclichydroxyalkyl, and R³ is substituted aryl.
 51. A compound according toclaim 41, wherein the R¹ group is the S-enantiomer.
 52. A pharmaceuticalcomposition comprising an effective amount of at least one compoundaccording to claim 41, and a pharmaceutically acceptable carrier.
 53. Amethod of treating cancer in a mammal comprising administering atherapeutically effective amount of a medicament comprising at least onecompound according to claim
 41. 54. A method according to claim 53,wherein the cancer overexpresses MDM2.
 55. A method according to claim53, wherein the cancer is selected from osteosarcoma, colorectalcarcinoma, neuroblastoma and uterus chorion cancer.
 56. A method ofinhibiting the interaction of MDM2 protein with p53 comprisingadministering a therapeutically effective amount of a compound accordingto claim 41 or a pharmaceutically acceptable salt thereof.
 57. A methodaccording to claim 53 further comprising administering a therapeuticallyeffective amount of at least one additional anticancer agent.
 58. Amethod according to claim 54 further comprising administering atherapeutically effective amount of at least one additional anticanceragent.
 59. A kit comprising at least one compound according to claim 41;and instructions for use.
 60. A method of manufacturing a medicamentcomprising, combining a compound according to claim 41 or apharmaceutically acceptable salt thereof, with a carrier that comprisesone or more accessory ingredients.